Annals of Biomedical Engineering

, Volume 42, Issue 10, pp 2014–2028 | Cite as

Asymmetric Cell–Matrix and Biomechanical Abnormalities in Elastin Insufficiency Induced Aortopathy

  • Varun K. Krishnamurthy
  • Ashlie N. Evans
  • Janaka P. Wansapura
  • Hanna Osinska
  • Kelsey E. Maddy
  • Stefanie V. Biechler
  • Daria A. Narmoneva
  • Richard L. Goodwin
  • Robert B. HintonEmail author


Aortopathy is characterized by vascular smooth muscle cell (VSMC) abnormalities and elastic fiber fragmentation. Elastin insufficient (Eln +/− ) mice demonstrate latent aortopathy similar to human disease. We hypothesized that aortopathy manifests primarily in the aorto-pulmonary septal (APS) side of the thoracic aorta due to asymmetric cardiac neural crest (CNC) distribution. Anatomic (aortic root vs. ascending aorta) and molecular (APS vs. non-APS) regions of proximal aorta tissue were examined in adult and aged wild type (WT) and mutant (Eln +/− ) mice. CNC, VSMCs, elastic fiber architecture, proteoglycan expression, morphometrics and biomechanical properties were examined using histology, 3D reconstruction, micropipette aspiration and in vivo magnetic resonance imaging (MRI). In the APS side of Eln +/− aorta, Sonic Hedgehog (SHH) is decreased while SM22 is increased. Elastic fiber architecture abnormalities are present in the Eln +/− aortic root and APS ascending aorta, and biglycan is increased in the aortic root while aggrecan is increased in the APS aorta. The Eln +/− ascending aorta is stiffer than the aortic root, the APS side is thicker and stiffer than the non-APS side, and significant differences in the individual aortic root sinuses are observed. Asymmetric structure–function abnormalities implicate regional CNC dysregulation in the development and progression of aortopathy.


Aortic root Cardiac neural crest Elastic fibers Micropipette aspiration Biomedical engineering 



Aorto-pulmonary septum


Cardiac neural crest


Extracellular matrix


Elastic fiber fragmentation


Homozygous deletion of elastin gene


Heterozygous deletion of elastin gene


Integrated optical density


Magnetic resonance imaging


Maximum rate of systolic distension


Persistent truncus arteriosus


Sonic hedgehog


Thoracic aortic aneurysm


Vascular smooth muscle cell


Wild type



We thank Amy Opoka for her assistance. We also thank Dr. Dean Y. Li (University of Utah) for providing the elastin knockout mice and Dr. Robert P. Mecham (Washington University) for helpful discussions. Present address for Varun K. Krishnamurthy: Department of Bioengineering, Rice University, Houston, TX 77005, USA. This work was supported by the AHA 11PRE7210044 (VKK), NIH HL086856-01 (RLG), NIH HL085122 (RBH), and Cincinnati Children’s Research Foundation (RBH).


The authors have nothing to disclose.

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  1. 1.
    Anderson, R. H. Clinical anatomy of the aortic root. Heart 84:670–673, 2000.PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Bauer, M., M. Pasic, R. Meyer, N. Goetze, U. Bauer, H. Siniawski, and R. Hetzer. Morphometric analysis of aortic media in patients with bicuspid and tricuspid aortic valve. Ann. Thorac. Surg. 74:58–62, 2002.PubMedCrossRefGoogle Scholar
  3. 3.
    Becker, A. E., M. J. Becker, and J. E. Edwards. Pathology of the semilunar valve in persistent truncus arteriosus. J. Thorac. Cardiovasc. Surg. 62:16–26, 1971.PubMedGoogle Scholar
  4. 4.
    Bergwerff, M., M. C. DeRuiter, R. E. Poelmann, and A. C. Gittenberger-de Groot. Onset of elastogenesis and downregulation of smooth muscle actin as distinguishing phenomena in artery differentiation in the chick embryo. Anat. Embryol. (Berl). 194:545–557, 1996.Google Scholar
  5. 5.
    Bergwerff, M., M. E. Verberne, M. C. DeRuiter, R. E. Poelmann, and A. C. Gittenberger-de Groot. Neural crest cell contribution to the developing circulatory system: implications for vascular morphology? Circ. Res. 82:221–231, 1998.PubMedCrossRefGoogle Scholar
  6. 6.
    Bruneau, B. G. The developmental genetics of congenital heart disease. Nature 451:943–948, 2008.PubMedCrossRefGoogle Scholar
  7. 7.
    Calder, L., R. Van Praagh, S. Van Praagh, W. P. Sears, R. Corwin, A. Levy, J. D. Keith, and M. H. Paul. Truncus arteriosus communis. Clinical, angiocardiographic, and pathologic findings in 100 patients. Am. Heart J. 92:23–38, 1976.PubMedCrossRefGoogle Scholar
  8. 8.
    Carlo, W. F., E. D. McKenzie, and T. C. Slesnick. Root dilation in patients with truncus arteriosus. Congenit. Heart Dis. 6:228–233, 2011.PubMedCrossRefGoogle Scholar
  9. 9.
    Cattell, M. A., P. S. Hasleton, and J. C. Anderson. Glycosaminoglycan content is increased in dissecting aneurysms of human thoracic aorta. Clin. Chim. Acta 226:29–46, 1994.PubMedCrossRefGoogle Scholar
  10. 10.
    Cheung, C., A. S. Bernardo, M. W. Trotter, R. A. Pedersen, and S. Sinha. Generation of human vascular smooth muscle subtypes provides insight into embryological origin-dependent disease susceptibility. Nat. Biotechnol. 30:165–173, 2012.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Choudhary, B., Y. Ito, T. Makita, T. Sasaki, Y. Chai, and H. M. Sucov. Cardiovascular malformations with normal smooth muscle differentiation in neural crest-specific type II TGFbeta receptor (Tgfbr2) mutant mice. Dev. Biol. 289:420–429, 2006.PubMedCrossRefGoogle Scholar
  12. 12.
    Creazzo, T. L., R. E. Godt, L. Leatherbury, S. J. Conway, and M. L. Kirby. Role of cardiac neural crest cells in cardiovascular development. Annu. Rev. Physiol. 60:267–286, 1998.PubMedCrossRefGoogle Scholar
  13. 13.
    Dodou, E., M. P. Verzi, J. P. Anderson, S. M. Xu, and B. L. Black. Mef2c is a direct transcriptional target of ISL1 and GATA factors in the anterior heart field during mouse embryonic development. Development 131:3931–3942, 2004.PubMedCrossRefGoogle Scholar
  14. 14.
    Donato Aquaro, G., L. Ait-Ali, M. L. Basso, M. Lombardi, A. Pingitore, and P. Festa. Elastic properties of aortic wall in patients with bicuspid aortic valve by magnetic resonance imaging. Am. J. Cardiol. 108:81–87, 2011.PubMedCrossRefGoogle Scholar
  15. 15.
    Eronen, M., M. Peippo, A. Hiippala, M. Raatikka, M. Arvio, R. Johansson, and M. Kahkonen. Cardiovascular manifestations in 75 patients with Williams syndrome. J. Med. Genet. 39:554–558, 2002.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Faury, G., M. Pezet, R. H. Knutsen, W. A. Boyle, S. P. Heximer, S. E. McLean, R. K. Minkes, K. J. Blumer, A. Kovacs, D. P. Kelly, D. Y. Li, B. Starcher, and R. P. Mecham. Developmental adaptation of the mouse cardiovascular system to elastin haploinsufficiency. J. Clin. Invest. 112:1419–1428, 2003.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Feng, M., S. Whitesall, Y. Zhang, M. Beibel, L. D’Alecy, and K. DiPetrillo. Validation of volume-pressure recording tail-cuff blood pressure measurements. Am. J. Hypertens. 21:1288–1291, 2008.PubMedCrossRefGoogle Scholar
  18. 18.
    Goddeeris, M. M., R. Schwartz, J. Klingensmith, and E. N. Meyers. Independent requirements for Hedgehog signaling by both the anterior heart field and neural crest cells for outflow tract development. Development 134:1593–1604, 2007.PubMedCrossRefGoogle Scholar
  19. 19.
    Goergen, C. J., K. N. Barr, D. T. Huynh, J. R. Eastham-Anderson, G. Choi, M. Hedehus, R. L. Dalman, A. J. Connolly, C. A. Taylor, P. S. Tsao, and J. M. Greve. In vivo quantification of murine aortic cyclic strain, motion, and curvature: implications for abdominal aortic aneurysm growth. J. Magn. Reson. Imaging 32:847–858, 2010.PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Grande, K. J., R. P. Cochran, P. G. Reinhall, and K. S. Kunzelman. Stress variations in the human aortic root and valve: the role of anatomic asymmetry. Ann. Biomed. Eng. 26:534–545, 1998.PubMedCrossRefGoogle Scholar
  21. 21.
    Guilak, F., L. G. Alexopoulos, M. A. Haider, H. P. Ting-Beall, and L. A. Setton. Zonal uniformity in mechanical properties of the chondrocyte pericellular matrix: micropipette aspiration of canine chondrons isolated by cartilage homogenization. Ann. Biomed. Eng. 33:1312–1318, 2005.PubMedCrossRefGoogle Scholar
  22. 22.
    Gundiah, N., K. Kam, P. B. Matthews, J. Guccione, H. A. Dwyer, D. Saloner, T. A. Chuter, T. S. Guy, M. B. Ratcliffe, and E. E. Tseng. Asymmetric mechanical properties of porcine aortic sinuses. Ann. Thorac. Surg. 85:1631–1638, 2008.PubMedCrossRefGoogle Scholar
  23. 23.
    Hahn, R. T., M. J. Roman, A. H. Mogtader, and R. B. Devereux. Association of aortic dilation with regurgitant, stenotic and functionally normal bicuspid aortic valves. J. Am. Coll. Cardiol. 19:283–288, 1992.PubMedCrossRefGoogle Scholar
  24. 24.
    Hallidie-Smith, K. A., and S. Karas. Cardiac anomalies in Williams-Beuren syndrome. Arch. Dis. Child. 63:809–813, 1988.PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Heiberg, E., J. Sjogren, M. Ugander, M. Carlsson, H. Engblom, and H. Arheden. Design and validation of segment—freely available software for cardiovascular image analysis. BMC Med. Imaging 10:1, 2010.PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Hinton, R. B. Bicuspid aortic valve and thoracic aortic aneurysm: three patient populations, two disease phenotypes, and one shared genotype. Cardiol. Res. Pract. 2012:926975, 2012.PubMedCentralPubMedGoogle Scholar
  27. 27.
    Hinton, R. B., J. Adelman-Brown, S. Witt, V. K. Krishnamurthy, H. Osinska, B. Sakthivel, J. F. James, D. Y. Li, D. A. Narmoneva, R. P. Mecham, and D. W. Benson. Elastin haploinsufficiency results in progressive aortic valve malformation and latent valve disease in a mouse model. Circ. Res. 107:549–557, 2010.PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Hinton, Jr., R. B., J. Lincoln, G. H. Deutsch, H. Osinska, P. B. Manning, D. W. Benson, and K. E. Yutzey. Extracellular matrix remodeling and organization in developing and diseased aortic valves. Circ. Res. 98:1431–1438, 2006.PubMedCrossRefGoogle Scholar
  29. 29.
    Hiratzka, L. F., G. L. Bakris, J. A. Beckman, R. M. Bersin, V. F. Carr, D. E. Casey, Jr., K. A. Eagle, L. K. Hermann, E. M. Isselbacher, E. A. Kazerooni, N. T. Kouchoukos, B. W. Lytle, D. M. Milewicz, D. L. Reich, S. Sen, J. A. Shinn, L. G. Svensson, and D. M. Williams. 2010 ACCF/AHA/AATS/ACR/ASA/SCA/SCAI/SIR/STS/SVM guidelines for the diagnosis and management of patients with Thoracic Aortic Disease: a report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines, American Association for Thoracic Surgery, American College of Radiology, American Stroke Association, Society of Cardiovascular Anesthesiologists, Society for Cardiovascular Angiography and Interventions, Society of Interventional Radiology, Society of Thoracic Surgeons, and Society for Vascular Medicine. Circulation 121:e266–e369, 2010.PubMedCrossRefGoogle Scholar
  30. 30.
    Ho, S. Y. Structure and anatomy of the aortic root. Eur. J. Echocardiogr. 10:i3–i10, 2009.PubMedCrossRefGoogle Scholar
  31. 31.
    Keane, J. F., K. E. Fellows, C. G. LaFarge, A. S. Nadas, and W. F. Bernhard. The surgical management of discrete and diffuse supravalvar aortic stenosis. Circulation 54:112–117, 1976.PubMedCrossRefGoogle Scholar
  32. 32.
    Kirby, M. L., T. F. Gale, and D. E. Stewart. Neural crest cells contribute to normal aorticopulmonary septation. Science 220:1059–1061, 1983.PubMedCrossRefGoogle Scholar
  33. 33.
    Kirby, M. L., and K. L. Waldo. Role of neural crest in congenital heart disease. Circulation 82:332–340, 1990.PubMedCrossRefGoogle Scholar
  34. 34.
    Krishnamurthy, V. K., F. Guilak, D. A. Narmoneva, and R. B. Hinton. Regional structure-function relationships in mouse aortic valve tissue. J. Biomech. 44:77–83, 2011.PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Krishnamurthy, V. K., A. M. Opoka, C. B. Kern, F. Guilak, D. A. Narmoneva, and R. B. Hinton. Maladaptive matrix remodeling and regional biomechanical dysfunction in a mouse model of aortic valve disease. Matrix Biol. 31:197–205, 2012.PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Li, D. Y., B. Brooke, E. C. Davis, R. P. Mecham, L. K. Sorensen, B. B. Boak, E. Eichwald, and M. T. Keating. Elastin is an essential determinant of arterial morphogenesis. Nature 393:276–280, 1998.PubMedCrossRefGoogle Scholar
  37. 37.
    Li, D. Y., G. Faury, D. G. Taylor, E. C. Davis, W. A. Boyle, R. P. Mecham, P. Stenzel, B. Boak, and M. T. Keating. Novel arterial pathology in mice and humans hemizygous for elastin. J. Clin. Invest. 102:1783–1787, 1998.PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Majesky, M. W. Developmental basis of vascular smooth muscle diversity. Arterioscler. Thromb. Vasc. Biol. 27:1248–1258, 2007.PubMedCrossRefGoogle Scholar
  39. 39.
    McLean, M., and J. W. Prothero. Three-dimensional reconstruction from serial sections. V. Calibration of dimensional changes incurred during tissue preparation and data processing. Anal. Quant. Cytol. Histol. 13:269–278, 1991.PubMedGoogle Scholar
  40. 40.
    Morrison, T. M., G. Choi, C. K. Zarins, and C. A. Taylor. Circumferential and longitudinal cyclic strain of the human thoracic aorta: age-related changes. J. Vasc. Surg. 49:1029–1036, 2009.PubMedCentralPubMedCrossRefGoogle Scholar
  41. 41.
    Nakamura, T., M. C. Colbert, and J. Robbins. Neural crest cells retain multipotential characteristics in the developing valves and label the cardiac conduction system. Circ. Res. 98:1547–1554, 2006.PubMedCrossRefGoogle Scholar
  42. 42.
    Neeb, Z., J. D. Lajiness, E. Bolanis, and S. J. Conway. Cardiac outflow tract anomalies. Wiley Interdiscip. Rev. Dev. Biol. 2:499–530, 2013.PubMedCentralPubMedCrossRefGoogle Scholar
  43. 43.
    Owens, G. K., M. S. Kumar, and B. R. Wamhoff. Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol. Rev. 84:767–801, 2004.PubMedCrossRefGoogle Scholar
  44. 44.
    Pachulski, R. T., A. L. Weinberg, and K. L. Chan. Aortic aneurysm in patients with functionally normal or minimally stenotic bicuspid aortic valve. Am. J. Cardiol. 67:781–782, 1991.PubMedCrossRefGoogle Scholar
  45. 45.
    Passman, J. N., X. R. Dong, S. P. Wu, C. T. Maguire, K. A. Hogan, V. L. Bautch, and M. W. Majesky. A sonic hedgehog signaling domain in the arterial adventitia supports resident Sca1+ smooth muscle progenitor cells. Proc. Natl Acad. Sci. U.S.A. 105:9349–9354, 2008.PubMedCentralPubMedCrossRefGoogle Scholar
  46. 46.
    Pezet, M., M. P. Jacob, B. Escoubet, D. Gheduzzi, E. Tillet, P. Perret, P. Huber, D. Quaglino, R. Vranckx, D. Y. Li, B. Starcher, W. A. Boyle, R. P. Mecham, and G. Faury. Elastin haploinsufficiency induces alternative aging processes in the aorta. Rejuvenation Res. 11:97–112, 2008.PubMedCentralPubMedCrossRefGoogle Scholar
  47. 47.
    Rosenquist, T. H., and A. C. Beall. Elastogenic cells in the developing cardiovascular system. Smooth muscle, nonmuscle, and cardiac neural crest. Ann. N. Y. Acad. Sci. 588:106–119, 1990.PubMedCrossRefGoogle Scholar
  48. 48.
    Rosenquist, T. H., A. C. Beall, L. Modis, and R. Fishman. Impaired elastic matrix development in the great arteries after ablation of the cardiac neural crest. Anat. Rec. 226:347–359, 1990.PubMedCrossRefGoogle Scholar
  49. 49.
    Schonherr, E., H. T. Jarvelainen, L. J. Sandell, and T. N. Wight. Effects of platelet-derived growth factor and transforming growth factor-beta 1 on the synthesis of a large versican-like chondroitin sulfate proteoglycan by arterial smooth muscle cells. J. Biol. Chem. 266:17640–17647, 1991.PubMedGoogle Scholar
  50. 50.
    Snarr, B. S., E. E. Wirrig, A. L. Phelps, T. C. Trusk, and A. Wessels. A spatiotemporal evaluation of the contribution of the dorsal mesenchymal protrusion to cardiac development. Dev. Dyn. 236:1287–1294, 2007.PubMedCentralPubMedCrossRefGoogle Scholar
  51. 51.
    Stoller, J. Z., and J. A. Epstein. Cardiac neural crest. Semin. Cell Dev. Biol. 16:704–715, 2005.PubMedCrossRefGoogle Scholar
  52. 52.
    Szabo, Z., M. W. Crepeau, A. L. Mitchell, M. J. Stephan, R. A. Puntel, K. Yin Loke, R. C. Kirk, and Z. Urban. Aortic aneurysmal disease and cutis laxa caused by defects in the elastin gene. J. Med. Genet. 43:255–258, 2006.PubMedCentralPubMedCrossRefGoogle Scholar
  53. 53.
    Tadros, T. M., M. D. Klein, and O. M. Shapira. Ascending aortic dilatation associated with bicuspid aortic valve: pathophysiology, molecular biology, and clinical implications. Circulation 119:880–890, 2009.PubMedCrossRefGoogle Scholar
  54. 54.
    Theret, D. P., M. J. Levesque, M. Sato, R. M. Nerem, and L. T. Wheeler. The application of a homogeneous half-space model in the analysis of endothelial cell micropipette measurements. J. Biomech. Eng. 110:190–199, 1988.PubMedCrossRefGoogle Scholar
  55. 55.
    Treasure, T., J. Pepper, T. Golesworthy, R. Mohiaddin, and R. H. Anderson. External aortic root support: NICE guidance. Heart 98:65–68, 2012.PubMedCrossRefGoogle Scholar
  56. 56.
    Wagenseil, J. E., and R. P. Mecham. Vascular extracellular matrix and arterial mechanics. Physiol. Rev. 89:957–989, 2009.PubMedCentralPubMedCrossRefGoogle Scholar
  57. 57.
    Wagenseil, J. E., N. L. Nerurkar, R. H. Knutsen, R. J. Okamoto, D. Y. Li, and R. P. Mecham. Effects of elastin haploinsufficiency on the mechanical behavior of mouse arteries. Am. J. Physiol. Heart Circ. Physiol. 289:H1209–H1217, 2005.PubMedCrossRefGoogle Scholar
  58. 58.
    Waldo, K. L., M. R. Hutson, C. C. Ward, M. Zdanowicz, H. A. Stadt, D. Kumiski, R. Abu-Issa, and M. L. Kirby. Secondary heart field contributes myocardium and smooth muscle to the arterial pole of the developing heart. Dev. Biol. 281:78–90, 2005.PubMedCrossRefGoogle Scholar
  59. 59.
    Wang, J., A. Nagy, J. Larsson, M. Dudas, H. M. Sucov, and V. Kaartinen. Defective ALK5 signaling in the neural crest leads to increased postmigratory neural crest cell apoptosis and severe outflow tract defects. BMC Dev. Biol. 6:51, 2006.PubMedCentralPubMedCrossRefGoogle Scholar
  60. 60.
    Washington Smoak, I., N. A. Byrd, R. Abu-Issa, M. M. Goddeeris, R. Anderson, J. Morris, K. Yamamura, J. Klingensmith, and E. N. Meyers. Sonic hedgehog is required for cardiac outflow tract and neural crest cell development. Dev. Biol. 283:357–372, 2005.PubMedCrossRefGoogle Scholar
  61. 61.
    Wyffels, J. T. Principles and techniques of electron microscopy: biological applications, fourth edition, by M. A. Hayat. Microsc. Microanal. 7:66, 2001.PubMedGoogle Scholar
  62. 62.
    Xie, W. B., Z. Li, N. Shi, X. Guo, J. Tang, W. Ju, J. Han, T. Liu, E. P. Bottinger, Y. Chai, P. A. Jose, and S. Y. Chen. Smad2 and myocardin-related transcription factor B cooperatively regulate vascular smooth muscle differentiation from neural crest cells. Circ. Res. 113:e76–e86, 2013.PubMedCrossRefGoogle Scholar
  63. 63.
    Ke, Y., and R. Sukthankar. PCA-SIFT: a more distinctive representation for local image descriptors. Proceedings of Computer Vision and Pattern Recognition, 2004.Google Scholar
  64. 64.
    Yacoub, M. H., P. J. Kilner, E. J. Birks, and M. Misfeld. The aortic outflow and root: a tale of dynamism and crosstalk. Ann. Thorac. Surg. 68:S37–S43, 1999.PubMedCrossRefGoogle Scholar
  65. 65.
    Zhao, X., R. Pratt, and J. Wansapura. Quantification of aortic compliance in mice using radial phase contrast MRI. J. Magn. Reson. Imaging 30:286–291, 2009.PubMedCrossRefGoogle Scholar

Copyright information

© Biomedical Engineering Society 2014

Authors and Affiliations

  • Varun K. Krishnamurthy
    • 1
    • 2
  • Ashlie N. Evans
    • 3
  • Janaka P. Wansapura
    • 4
  • Hanna Osinska
    • 5
  • Kelsey E. Maddy
    • 6
  • Stefanie V. Biechler
    • 6
  • Daria A. Narmoneva
    • 2
  • Richard L. Goodwin
    • 3
  • Robert B. Hinton
    • 1
    Email author
  1. 1.Division of Cardiology, the Heart InstituteCincinnati Children’s Hospital Medical CenterCincinnatiUSA
  2. 2.Department of Biomedical EngineeringUniversity of CincinnatiCincinnatiUSA
  3. 3.Department of Biomedical EngineeringUniversity of South CarolinaColumbiaUSA
  4. 4.Division of RadiologyCincinnati Children’s Hospital Medical CenterCincinnatiUSA
  5. 5.Division of Molecular Cardiovascular BiologyCincinnati Children’s Hospital Medical CenterCincinnatiUSA
  6. 6.Department of Cell Biology and AnatomyUniversity of South CarolinaColumbiaUSA

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