Advertisement

Current Medical Science

, Volume 38, Issue 5, pp 765–775 | Cite as

Role of TGF-β1 Signaling in Heart Valve Calcification Induced by Abnormal Mechanical Stimulation in a Tissue Engineering Model

  • Xing-jian Hu
  • Wen-cong-hui Wu
  • Nian-guo DongEmail author
  • Jia-wei Shi
  • Jun-wei Liu
  • Si Chen
  • Chen Deng
  • Feng Shi
Article
  • 45 Downloads

Abstract

A tissue engineering model of heart valve calcification induced in a bioreactor was established to evaluate the calcification induced by abnormal mechanical stimulation and explore the underlying molecular mechanisms. Polyethylene glycol (PEG)-modified decellularized porcine aortic leaflets seeded with human valve interstitial cells (huVICs) were mounted on a Ti-Ni alloy frame to fabricate two-leaflet and threeleaflet tissue engineered valves. The two-leaflet model valves were exposed to abnormal pulsatile flow stimulation with null (group A), low (1000 mL/min, group B), medium (2000 mL/min, group C), and high velocity (3000 mL/min, group D) for 14 days. Morphology and calcification were assessed by von Kossa staining, alkaline phosphatase (ALP) content, and Runx2 immunostaining. Leaflet calcification and mRNA and protein expression of transforming growth factor (TGF)-β1, bone morphogenetic protein 2 (BMP2), Smad1, and MSX2 were measured at different time points. ALP content was examined in two-leaflet valves seeded with BMP2 shRNA plasmid-infected huVICs and exposed to the same stimulation conditions. The results showed that during 14 days of flow stimulation, huVICs on the leaflet surface proliferated to generate normal monolayer coverage in groups A, B, and C. Under mechanical stimulation, huVICs showed a parallel growth pattern in the direction of the fluid flow, but huVICs exhibited disordered growth in the high-velocity flow environment. von Kossa staining, ALP measurement, and immunohistochemical staining for Runx2 confirmed the lack of obvious calcification in group A and significant calcification in group D. Expression levels of TGF-β1, BMP2, and MSX2 mRNA and protein were increased under fluid stimulation. ALP production by BMP2 shRNA plasmid-infected huVICs on model leaflets was significantly reduced. In conclusion, abnormal mechanical stimulation in a bioreactor induced calcification in the tissue engineering valve model. The extent of calcification correlated positively with the flow velocity, as did the mRNA and protein levels of TGF-β1, BMP2, and MSX2. These findings indicate that TGF-β1/BMP2 signaling is involved in valve calcification induced by abnormal mechanical stimulation.

Key words

valve calcification abnormal mechanical stimulation bioreactor TGF-β1 signal pathway 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Hu SS, Kong LZ. Report on cardiovascular disease report in China (2011). Beijing: Encyclopedia of China Publishing House, 2012,1.Google Scholar
  2. 2.
    Nkomo VT, Gardin JM, Skelton TN, et al. Burden of valvular heart diseases: a population-based study. Lancet, 2006,368(9540):1005–1011CrossRefGoogle Scholar
  3. 3.
    Koyama J. Prognostic significance of occult transthyretin cardiac amyloidosis in patients with severe aortic stenosis undergoing surgical aortic valve replacement: An unrecognized disease modifier. Circ Cardiovasc Imaging, 2016,9(8):e005320CrossRefGoogle Scholar
  4. 4.
    Braun J, Klautz RJ. Mitral valve surgery in low ejection fraction, severe ischemic mitral regurgitation patients: should we repair them all? Curr Opin Cardiol, 2012,27(2):111–117CrossRefGoogle Scholar
  5. 5.
    Ustunsoy H, Gokaslan G, Ozcaliskan O, et al. "V-PLASTY": a novel technique to reconstruct pulmonary valvular and annular stenosis in patients with right ventricular outflow tract obstruction. J Cardiothorac Surg, 2013,8(3):55CrossRefGoogle Scholar
  6. 6.
    Brennan JM, Edwards FH, Zhao Y, et al. O. Longterm safety and effectiveness of mechanical versus biologic aortic valve prostheses in older patients: results from the Society of Thoracic Surgeons Adult Cardiac Surgery National Database. Circulation, 2013,127(16):1647–1655CrossRefGoogle Scholar
  7. 7.
    Yetkin E, Waltenberger J. Molecular and cellular mechanisms of aortic stenosis. Int J Cardiol, 2009,135(1):4–13CrossRefGoogle Scholar
  8. 8.
    Hakuno D, Kimura N, Yoshioka M, et al. Molecular mechanisms underlying the onset of degenerative aortic valve disease. J Mol Med (Berl), 2009,87(1):17–24CrossRefGoogle Scholar
  9. 9.
    Helske S, Kupari M, Lindstedt KA, et al. Aortic valve stenosis: an active atheroinflammatory process. Curr Opin Lipidol, 2007,18(5):483–491CrossRefGoogle Scholar
  10. 10.
    Miller JD, Weiss RM, Heistad DD. Calcific aortic valve stenosis: methods, models, and mechanisms. Circ Res, 2011,108(11):1392–1412CrossRefGoogle Scholar
  11. 11.
    Arjunon S, Rathan S, Jo H, et al. Aortic valve: mechanical environment and mechanobiology. Ann Biomed Eng, 2013,41(7):1331–1346CrossRefGoogle Scholar
  12. 12.
    Balachandran K, Sucosky P, Yoganathan AP. Hemodynamics and mechanobiology of aortic valve inflammation and calcification. Int J Inflam, 2011,2011(1):263870Google Scholar
  13. 13.
    Wipff PJ, Rifkin DB, Meister JJ, et al. Myofibroblast contraction activates latent TGF-1 from the extracellular matrix. J Cell Biol, 2007,179(6):1311–1323CrossRefGoogle Scholar
  14. 14.
    Kokubo H, Miyagawa-Tomita S, Tomimatsu H, et al. Targeted disruption of hesr2 results in atrioventricular valve anomalies that lead to heart dysfunction. Circ Res, 2004,95(5):540–547CrossRefGoogle Scholar
  15. 15.
    Guerraty M, Mohler Iii ER. Models of aortic valve calcification. J Investig Med, 2007,55(6):278–83CrossRefGoogle Scholar
  16. 16.
    Hu XJ, Dong NG, Shi JW, et al. Evaluation of a novel tetra-functional branched poly(ethylene glycol) crosslinker for manufacture of crosslinked, decellularized, porcine aortic valve leaflets. J Biomed Mater Res B Appl Biomater, 2014,102(2):322–326CrossRefGoogle Scholar
  17. 17.
    Hu XJ, Dong NG, Shi JW, et al. Synthesis and applications of tetra-functional branched poly-(ethylene glycol) derivative for the decellularized valve leaflets cross-linking. J Wuhan Univ Tech: Mater Sci, 2015,30(1):193–197CrossRefGoogle Scholar
  18. 18.
    Syedain ZH, Tranquillo RT. Controlled cyclic stretch bioreactor for tissue-engineered heart valves. Biomaterials, 2009,30(25):4078–4084CrossRefGoogle Scholar
  19. 19.
    Engelmayr GC Jr, Soletti L, Vigmostad SC, et al. A novel flex-stretch-flow bioreactor for the study of engineered heart valve tissue mechanobiology. Ann Biomed Eng, 2008,36(5):700–712CrossRefGoogle Scholar
  20. 20.
    Sage AP, Tintut Y, Demer LL. Regulatory mechanisms in vascular calcification. Nat Rev Cardiol, 2010,7(9):528–536CrossRefGoogle Scholar
  21. 21.
    Jian B, Narula N, Li QY, et al. Progression of aortic valve stenosis: TGF-beta1 is present in calcified aortic valve cusps and promotes aortic valve interstitial cell calcification via apoptosis. Ann Thorac Surg, 2003,75(2):457–465; discussion 465–466CrossRefGoogle Scholar
  22. 22.
    Cao X, Chen D. The BMP signaling and in vivo bone formation. Gene, 2005,357(1):1–8CrossRefGoogle Scholar
  23. 23.
    Kaden JJ, Bickelhaupt S, Grobholz R, et al. Expression of bone sialoprotein and bone morphogenetic protein-2 in calcific aortic stenosis. J Heart Valve Dis, 2004,13(4):560–566Google Scholar
  24. 24.
    Balachandran K, Sucosky P, Jo H, et al. Elevated cyclic stretch induces aortic valve calcification in a bone morphogenic protein-dependent manner. Am J Pathol, 2010,177(1):49–57CrossRefGoogle Scholar
  25. 25.
    Deng C, Dong N, Shi J, et al. Application of decellularized scaffold combined with loaded nanoparticles for heart valve tissue engineering in vitro. J Huazhong Univ Sci Technolog Med Sci, 2011,31(1):88–93CrossRefGoogle Scholar
  26. 26.
    Chester AH, Taylor PM. Molecular and functional characteristics of heart-valve interstitial cells. Philos Trans R Soc Lond B Biol Sci, 2007,362(1484):1437–1443CrossRefGoogle Scholar
  27. 27.
    Bae WJ, Lee SH, Rho YS, et al. Transforming growth factor β1 enhances stemness of head and neck squamous cell carcinoma cells through activation of Wnt signaling. Oncol Lett, 2016,12(6):5315–5320CrossRefGoogle Scholar
  28. 28.
    Wang W, Koka V, Lan HY. Transforming growth factor-beta and Smad signalling in kidney diseases. Nephrology (Carlton), 2005,10(1):48–56CrossRefGoogle Scholar
  29. 29.
    Merryman WD, Lukoff HD, Long RA, et al. Synergistic effects of cyclic tension and transforming growth factor beta1 on the aortic valve myofibroblast. Card Path, 2007,16(5):268–276CrossRefGoogle Scholar
  30. 30.
    Retting KN, Song B, Yoon BS, et al. BMP canonical Smad signaling through Smad1 and Smad5 is required for endochondral bone formation. Development, 2009,136(7):1093–1104CrossRefGoogle Scholar
  31. 31.
    Song B, Estrada KD, Lyons KM. Smad signaling in skeletal development and regeneration. Cytokine Growth Factor Rev, 2009,20(5–6):379–388CrossRefGoogle Scholar
  32. 32.
    Massagué J, Seoane J, Wotton D. Smad transcription factors. Genes Dev, 2005,19(23):2783–2810CrossRefGoogle Scholar
  33. 33.
    Shao JS, Cheng SL, Pingsterhaus JM, et al. Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals. J Clin Invest, 2005,115(5):1210–1220CrossRefGoogle Scholar
  34. 34.
    Reilly GC, Golden EB, Grasso-Knight G, et al. Differential effects of ERK and p38 signaling in BMP-2 stimulated hypertrophy of cultured chick sternal chondrocytes. Cell Commun Signal, 2005,3(1):3–10CrossRefGoogle Scholar

Copyright information

© Huazhong University of Science and Technology 2018

Authors and Affiliations

  • Xing-jian Hu
    • 1
  • Wen-cong-hui Wu
    • 2
  • Nian-guo Dong
    • 1
  • Jia-wei Shi
    • 1
  • Jun-wei Liu
    • 1
  • Si Chen
    • 1
  • Chen Deng
    • 1
  • Feng Shi
    • 1
  1. 1.Department of Cardiovascular Surgery, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Department of GastroenterologyZhongnan Hospital of Wuhan UniversityWuhanChina

Personalised recommendations