Skip to main content
SpringerLink
Log in

Relative Effects of Fluid Oscillations and Nutrient Transport in the In Vitro Growth of Valvular Tissues

  • Published:
Cardiovascular Engineering and Technology Aims and scope Submit manuscript

Abstract

Engineered valvular tissues are cultured dynamically, and involve specimen movement. We previously demonstrated that oscillatory shear stresses (OSS) under combined steady flow and specimen cyclic flexure (flex-flow) promote tissue formation. However, localized efficiency of specimen mass transport is also important in the context of cell viability within the growing tissues. Here, we investigated the delivery of two essential species for cell survival, glucose and oxygen, to 3-dimensional (3D) engineered valvular tissues. We applied a convective-diffusive model to characterize glucose and oxygen mass transport with and without valve-like specimen flexural movement. We found the mass transport effects for glucose and oxygen to be negligible for scaffold porosities typically present during in vitro experiments and non-essential unless the porosity was unusually low (<40%). For more typical scaffold porosities (75%) however, we found negligible variation in the specimen mass fraction of glucose and oxygen in both non-moving and moving constructs (p > 0.05). Based on this result, we conducted an experiment using bone marrow stem cell (BMSC)-seeded scaffolds under Pulsatile flow-alone states to permit OSS without any specimen movement. BMSC-seeded specimen collagen from the pulsatile flow and flex-flow environments were subsequently found to be comparable (p > 0.05) and exhibited some gene expression similarities. We conclude that a critical magnitude of fluid-induced, OSS created by either pulsatile flow or flex-flow conditions, particularly when the oscillations are physiologically-relevant, is the direct, principal stimulus that promotes engineered valvular tissues and its phenotype, whereas mass transport benefits derived from specimen movement are minimal.

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.

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Abraham, J., et al. A mass transfer model of temporal drug deposition in artery walls. Int. J. Heat Mass Transf. 58(1):632–638, 2013.

    Article  Google Scholar 

  2. Ai, L., and K. Vafai. A coupling model for macromolecule transport in a stenosed arterial wall. Int. J. Heat Mass Transf. 49(9):1568–1591, 2006.

    Article  MATH  Google Scholar 

  3. Boubriak, O., J. Urban, and Z. Cui. Monitoring of lactate and glucose levels in engineered cartilage construct by microdialysis. J. Membr. Sci. 273(1):77–83, 2006.

    Article  Google Scholar 

  4. Brown, D. A., et al. Analysis of oxygen transport in a diffusion-limited model of engineered heart tissue. Biotechnol. Bioeng. 97(4):962–975, 2007.

    Article  Google Scholar 

  5. Chen, G.-Q., and Q. Wu. The application of polyhydroxyalkanoates as tissue engineering materials. Biomaterials 26(33):6565–6578, 2005.

    Article  Google Scholar 

  6. Chen, J., et al. Numerical investigation of mass transport through patient-specific deformed aortae. J. Biomech. 47(2):544–552, 2014.

    Article  Google Scholar 

  7. Chung, S., and K. Vafai. Low-density lipoprotein transport within a multi-layered arterial wall—effect of the atherosclerotic plaque/stenosis. J. Biomech. 46(3):574–585, 2013.

    Article  Google Scholar 

  8. Chung, S., and K. Vafai. Mechanobiology of low-density lipoprotein transport within an arterial wall—Impact of hyperthermia and coupling effects. J. Biomech. 47(1):137–147, 2014.

    Article  Google Scholar 

  9. Converti, A., et al. Evaluation of glucose diffusion coefficient through cell layers for the kinetic study of an immobilized cell bioreactor. Chem. Eng. Sci. 51(7):1023–1026, 1996.

    Article  Google Scholar 

  10. Das, D. B. Multiscale simulation of nutrient transport in hollow fibre membrane bioreactor for growing bone tissue: sub-cellular scale and beyond. Chem. Eng. Sci. 62(13):3627–3639, 2007.

    Article  Google Scholar 

  11. Day, P., et al. What factors determine placental glucose transfer kinetics? Placenta 34(10):953–958, 2013.

    Article  Google Scholar 

  12. De Monte, F., G. Pontrelli, and S. Becker. Drug release in biological tissues. Transport in biological media. Amsterdam: Elsevier Science BV, 2013.

    Google Scholar 

  13. Dohmen, P. M., et al. Ten years of clinical results with a tissue-engineered pulmonary valve. Ann. Thorac. Surg. 92(4):1308–1314, 2011.

    Article  Google Scholar 

  14. Engelmayr, G. C., et al. A novel bioreactor for the dynamic flexural stimulation of tissue engineered heart valve biomaterials. Biomaterials 24(14):2523–2532, 2003.

    Article  Google Scholar 

  15. Engelmayr, G. C., et al. Cyclic flexure and laminar flow synergistically accelerate mesenchymal stem cell-mediated engineered tissue formation: implications for engineered heart valve tissues. Biomaterials 27(36):6083–6095, 2006.

    Article  Google Scholar 

  16. Engelmayr, Jr, G. C., et al. A novel flex-stretch-flow bioreactor for the study of engineered heart valve tissue mechanobiology. Ann. Biomed. Eng. 36(5):700–712, 2008.

    Article  Google Scholar 

  17. Fong, P., et al. The use of polymer based scaffolds in tissue-engineered heart valves. Prog Pediatr Cardiol 21(2):193–199, 2006.

    Article  Google Scholar 

  18. Frank, B. S., et al. Determining cell seeding dosages for tissue engineering human pulmonary valves. J. Surg. Res. 174(1):39–47, 2012.

    Article  Google Scholar 

  19. Gao, Z., and C. W. Xu. Glucose metabolism induces mono-ubiquitination of histone H2B in mammalian cells. Biochem. Biophys. Res. Commun. 404(1):428–433, 2011.

    Article  Google Scholar 

  20. Gill, J., et al. Modeling oxygen transport in human placental terminal villi. J. Theor. Biol. 291:33–41, 2011.

    Article  Google Scholar 

  21. Hilfiker, A., et al. Mesenchymal stem cells and progenitor cells in connective tissue engineering and regenerative medicine: is there a future for transplantation? Langenbeck’s Arch. Surg. 396(4):489–497, 2011.

    Article  Google Scholar 

  22. Hoerstrup, S. P., et al. Fluorescence activated cell sorting: a reliable method in tissue engineering of a bioprosthetic heart valve. Ann. Thorac. Surg. 66(5):1653–1657, 1998.

    Article  Google Scholar 

  23. Hoerstrup, S. P., et al. Tissue engineering of a bioprosthetic heart valve: stimulation of extracellular matrix assessed by hydroxyproline assay. ASAIO J. 45(5):397, 1999.

    Article  Google Scholar 

  24. Hoerstrup, S. P., et al. Functional living trileaflet heart valves grown in vitro. Circulation 102(suppl 3):Iii-44–Iii-49, 2000.

    Google Scholar 

  25. Hoerstrup, S. P., et al. New pulsatile bioreactor for in vitro formation of tissue engineered heart valves. Tissue Eng. 6(1):75–79, 2000.

    Article  Google Scholar 

  26. Hoffman, J. Incidence of congenital heart disease: I Postnatal incidence. Pediatr. Cardiol. 16(3):103–113, 1995.

    Article  Google Scholar 

  27. Ishaug-Riley, S. L., et al. Three-dimensional culture of rat calvarial osteoblasts in porous biodegradable polymers. Biomaterials 19(15):1405–1412, 1998.

    Article  Google Scholar 

  28. Johnson, A. S., et al. Oxygen consumption and diffusion in assemblages of respiring spheres: performance enhancement of a bioartificial pancreas. Chem. Eng. Sci. 64(22):4470–4487, 2009.

    Article  Google Scholar 

  29. Kaazempur-Mofrad, M., et al. Mass transport and fluid flow in stenotic arteries: axisymmetric and asymmetric models. Int. J. Heat Mass Transf. 48(21):4510–4517, 2005.

    Article  MATH  Google Scholar 

  30. Kadner, A., et al. A new source for cardiovascular tissue engineering: human bone marrow stromal cells. Eur. J. Cardiothorac. Surg. 21(6):1055–1060, 2002.

    Article  Google Scholar 

  31. Karner, G., and K. Perktold. Effect of endothelial injury and increased blood pressure on albumin accumulation in the arterial wall: a numerical study. J. Biomech. 33(6):709–715, 2000.

    Article  Google Scholar 

  32. Khakpour, M., and K. Vafai. Critical assessment of arterial transport models. Int. J. Heat Mass Transf. 51(3):807–822, 2008.

    Article  MATH  Google Scholar 

  33. Koutny, T. Estimating reaction delay for glucose level prediction. Med. Hypotheses 77(6):1034–1037, 2011.

    Article  Google Scholar 

  34. Kumar, R., F. Stepanek, and A. Mantalaris. An oxygen transport model for human bone marrow microcirculation. Food Bioprod. Process. 82(2):105–116, 2004.

    Article  Google Scholar 

  35. Leor, J., Y. Amsalem, and S. Cohen. Cells, scaffolds, and molecules for myocardial tissue engineering. Pharmacol. Ther. 105(2):151–163, 2005.

    Article  Google Scholar 

  36. Lichtenberg, A., et al. Cell seeded tissue engineered cardiac valves based on allograft and xenograft scaffolds. Prog. Pediatr. Cardiol. 21(2):211–217, 2006.

    Article  MathSciNet  Google Scholar 

  37. Liu, X., et al. Effect of non-Newtonian and pulsatile blood flow on mass transport in the human aorta. J. Biomech. 44(6):1123–1131, 2011.

    Article  Google Scholar 

  38. Lotz, J., et al. Cardiovascular flow measurement with phase-contrast MR imaging: basic facts and implementation 1. Radiographics 22(3):651–671, 2002.

    Article  MathSciNet  Google Scholar 

  39. Martineau, L. C. Large enhancement of skeletal muscle cell glucose uptake and suppression of hepatocyte glucose-6-phosphatase activity by weak uncouplers of oxidative phosphorylation. Biochim. Biophys. Acta. 1820(2):133–150, 2012.

    Article  Google Scholar 

  40. Martinez, C., et al. Periodontal ligament cells cultured under steady-flow environments demonstrate potential for use in heart valve tissue engineering. Tissue Eng. Part A 19(3–4):458–466, 2012.

    Google Scholar 

  41. Moore, J., and C. Ethier. Oxygen mass transfer calculations in large arteries. J. Biomech. Eng. 119(4):469–475, 1997.

    Article  Google Scholar 

  42. Neuenschwander, S., and S. P. Hoerstrup. Heart valve tissue engineering. Transpl. Immunol. 12(3):359–365, 2004.

    Article  Google Scholar 

  43. Patnaik, S. S., et al. Decellularized scaffolds: concepts, methodologies, and applications in cardiac tissue engineering and whole-organ regeneration. Tissue regeneration: where nanostructure meets biology. 77–124, 2013.

  44. Perry, T. E., et al. Bone marrow as a cell source for tissue engineering heart valves. Ann. Thorac. Surg. 75(3):761–767, 2003.

    Article  Google Scholar 

  45. Prosi, M., et al. Mathematical and numerical models for transfer of low-density lipoproteins through the arterial walls: a new methodology for the model set up with applications to the study of disturbed lumenal flow. J. Biomech. 38(4):903–917, 2005.

    Article  Google Scholar 

  46. Ramaswamy, S., et al. The role of organ level conditioning on the promotion of engineered heart valve tissue development in vitro using mesenchymal stem cells. Biomaterials 31(6):1114–1125, 2010.

    Article  MathSciNet  Google Scholar 

  47. Ramaswamy, S., et al. A novel bioreactor for mechanobiological studies of engineered heart valve tissue formation under pulmonary arterial physiological flow conditions. J. Biomech. Eng. 136(12):121009, 2014.

    Article  Google Scholar 

  48. Rath, S., et al. Differentiation and distribution of marrow stem cells in flex-flow environments demonstrate support of the valvular phenotype. PLoS ONE 10(11):e0141802, 2015.

    Article  MathSciNet  Google Scholar 

  49. Reddy, J., V. U. Unnikrishnan, and G. U. Unnikrishnan. Recent advances in the analysis of nanotube-reinforced polymeric biomaterials. J. Mech. Behav. Mater. 22(5–6):137–148, 2013.

    Google Scholar 

  50. Roger, V. L., et al. Heart disease and stroke statistics—2011 update a report from the American Heart Association. Circulation 123(4):e18–e209, 2011.

    Article  Google Scholar 

  51. Ruel, J., and G. Lachance. A new bioreactor for the development of tissue-engineered heart valves. Ann. Biomed. Eng. 37(4):674–681, 2009.

    Article  Google Scholar 

  52. Salinas, M., and S. Ramaswamy. Computational simulations predict a key role for oscillatory fluid shear stress in de novo valvular tissue formation. J. Biomech. 47(14):3517–3523, 2014.

    Article  Google Scholar 

  53. Salinas, M., et al. Oscillatory shear stress created by fluid pulsatility versus flexed specimen configurations. Comput. Method Biomech. Biomed. Eng. 17(7):728–739, 2014.

    Article  MathSciNet  Google Scholar 

  54. Schmidt, D., et al. Engineering of biologically active living heart valve leaflets using human umbilical cord-derived progenitor cells. Tissue Eng. 12(11):3223–3232, 2006.

    Article  Google Scholar 

  55. Schmidt, D., et al. Minimally-invasive implantation of living tissue engineered heart valves: a comprehensive approach from autologous vascular cells to stem cells. J. Am. Coll. Cardiol. 56(6):510–520, 2010.

    Article  Google Scholar 

  56. Sengers, B., et al. Computational study of culture conditions and nutrient supply in cartilage tissue engineering. Biotechnol. Prog. 21(4):1252–1261, 2005.

    Article  Google Scholar 

  57. Siepe, M., et al. Stem cells used for cardiovascular tissue engineering. Eur. J. Cardiothorac. Surg. 34(2):242–247, 2008.

    Article  Google Scholar 

  58. Simon, P., et al. Tissue Engineering of heart valves—Immunologic and inflammatory challenges of the allograft scaffold. Prog. Pediatr. Cardiol. 21(2):161–165, 2006.

    Article  Google Scholar 

  59. Sodian, R., et al. Tissue engineering of a trileaflet heart valve-early in vitro experiences with a combined polymer. Tissue Eng. 5(5):489, 1999.

    Article  Google Scholar 

  60. Sodian, R., et al. Early in vivo experience with tissue-engineered trileaflet heart valves. Circulation 102(19 Suppl 3):III22, 2000.

    Google Scholar 

  61. Sodian, R., et al. Tissue engineering of heart valves: in vitro experiences. Ann. Thorac. Surg. 70(1):140, 2000.

    Article  Google Scholar 

  62. Sodian, R., et al. Fabrication of a trileaflet heart valve scaffold from a polyhydroxyalkanoate biopolyester for use in tissue engineering. Tissue Eng. 6(2):183, 2000.

    Article  Google Scholar 

  63. Soukane, D. M., A. Shirazi-Adl, and J. Urban. Computation of coupled diffusion of oxygen, glucose and lactic acid in an intervertebral disc. J. Biomech. 40(12):2645–2654, 2007.

    Article  Google Scholar 

  64. Stangeby, D. K., and C. R. Ethier. Coupled computational analysis of arterial ldl transport—effects of hypertension. Comput. Method Biomech. Biomed. Eng. 5(3):233, 2002.

    Article  Google Scholar 

  65. Stella, J. A., J. Liao, and M. S. Sacks. Time-dependent biaxial mechanical behavior of the aortic heart valve leaflet. J. Biomech. 40(14):3169, 2007.

    Article  Google Scholar 

  66. Sutherland, F. W. H., et al. From stem cells to viable autologous semilunar heart valve. Circulation 111(21):2783, 2005.

    Article  Google Scholar 

  67. Tarbell, J. M., M. J. Lever, and C. G. Caro. The effect of varying albumin concentration of the hydraulic conductivity of the rabbit common carotid artery. Microvasc. Res. 35(2):204, 1988.

    Article  Google Scholar 

  68. Unnikrishnan, G. U., V. U. Unnikrishnan, and J. N. Reddy. Finite element model for nutrient distribution analysis of a hollow fiber membrane bioreactor. Int. J. Numer. Methods Biomed. Eng. 28(2):229, 2012.

    Article  MATH  Google Scholar 

  69. Vermot, J., et al. Reversing blood flows act through klf2a to ensure normal valvulogenesis in the developing heart (reversing blood flows during valvulogenesis). PLoS Biol. 7(11):e1000246, 2009.

    Article  Google Scholar 

  70. Wang, L., et al. Computational simulation of oxygen diffusion in aortic valve leaflet for tissue engineering applications. J. Heart valve Dis. 17(6):700, 2008.

    Google Scholar 

  71. Wang, L., et al. Factors influencing the oxygen consumption rate of aortic valve interstitial cells: application to tissue engineering (report). Tissue Eng. Part C Method 15(3):355, 2009.

    Article  Google Scholar 

  72. Wang, L., et al. Prediction of oxygen distribution in aortic valve leaflet considering diffusion and convection. J. Heart valve Dis. 20(4):442, 2011.

    Google Scholar 

  73. Weber, B., S. M. Zeisberger, and S. P. Hoerstrup. Prenatally harvested cells for cardiovascular tissue engineering: fabrication of autologous implants prior to birth. Placenta 32:S316, 2011.

    Article  Google Scholar 

  74. Weber, B., et al. Engineering of living autologous human umbilical cord cell-based septal occluder membranes using composite PGA-P4HB matrices. Biomaterials 32(36):9630, 2011.

    Article  Google Scholar 

  75. Yang, N., and K. Vafai. Modeling of low-density lipoprotein (LDL) transport in the artery—effects of hypertension. Int. J. Heat Mass Transf. 49(5):850, 2006.

    Article  MATH  Google Scholar 

  76. Yu, P., et al. Fluid dynamics and oxygen transport in a micro-bioreactor with a tissue engineering scaffold. Int. J. Heat Mass Transf. 52(1):316, 2009.

    Article  MATH  Google Scholar 

Download references

Acknowledgments

The authors acknowledge funds received from the department of Biomedical Engineering at Florida International University to carry out this research work. Funding for M.S. by NIH/NIGMS R25 GM061347 is gratefully acknowledged.

Conflict of interest

The authors have no conflict of interests.

Statement of Human and Animal Studies

No Human and Animal Studies were involved in this study.

Author information

Authors and Affiliations

  1. Tissue Engineering, Mechanics, Imaging, and Materials Laboratory, Department of Biomedical Engineering, College of Engineering and Computing, Florida International University, 10555 W. Flagler Street, EC 2612, Miami, FL, 33174, USA

    Manuel Salinas, Sasmita Rath, Ana Villegas & Sharan Ramaswamy

  2. Department of Aerospace Engineering and Mechanics, The University of Alabama, Tuscaloosa, AL, USA

    Vinu Unnikrishnan

Authors
  1. Manuel Salinas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sasmita Rath
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ana Villegas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vinu Unnikrishnan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sharan Ramaswamy
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sharan Ramaswamy.

Additional information

Associate Editor Ajit P. Yoganathan oversaw the review of this article.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Salinas, M., Rath, S., Villegas, A. et al. Relative Effects of Fluid Oscillations and Nutrient Transport in the In Vitro Growth of Valvular Tissues. Cardiovasc Eng Tech 7, 170–181 (2016). https://doi.org/10.1007/s13239-016-0258-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13239-016-0258-x

Keywords

Search

Navigation