Tissue engineered heart valves (TEHV) are being investigated to address the limitations of currently available valve prostheses. In order to advance a wide variety of TEHV approaches, the goal of this study was to develop a cardiac valve bioreactor system capable of conditioning living valves with a range of hydrodynamic conditions as well as capable of assessing hydrodynamic performance to ISO 5840 standards.
A bioreactor system was designed based on the Windkessel approach. Novel features including a purpose-built valve chamber and pressure feedback control were incorporated to maintain asepsis while achieving a range of hydrodynamic conditions. The system was validated by testing hydrodynamic conditions with a bioprosthesis and by operating with cell culture medium for 4 weeks and living cells for 2 weeks.
The bioreactor system was able to produce a range of pressure and flow conditions from static to resting adult left ventricular outflow tract to pathological including hypertension. The system operated aseptically for 4 weeks and cell viability was maintained for 2 weeks. The system was also able to record the pressure and flow data needed to calculate effective orifice area and regurgitant fraction.
We have developed a single bioreactor system that allows for step-wise conditioning protocols to be developed for each unique TEHV design as well as allows for hydrodynamic performance assessment.
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Aleksieva, G., T. Hollweck, N. Thierfelder, U. Haas, F. Koenig, C. Fano, M. Dauner, E. Wintermantel, B. Reichart, C. Schmitz, and B. Akra. Use of a special bioreactor for the cultivation of a new flexible polyurethane scaffold for aortic valve tissue engineering. Biomed. Eng. Online 11:92, 2012.
Amrollahi, P., and L. Tayebi. Bioreactors for heart valve tissue engineering: a review. J. Chem. Technol. Biotechnol. 91:847–856, 2016.
Barzilla, J. E., A. S. McKenney, A. E. Cowan, C. A. Durst, and K. J. Grande-Allen. Design and validation of a novel splashing bioreactor system for use in mitral valve organ culture. Ann. Biomed. Eng. 38:3280–3294, 2010.
Benjamin, E. J., M. J. Blaha, S. E. Chiuve, M. Cushman, S. R. Das, R. Deo, S. D. deFerranti, J. Floyd, M. Fornage, C. Gillespie, C. R. Isasi, M. C. Jimenez, L. C. Jordan, S. E. Judd, D. Lackland, J. H. Lichtman, L. Lisabeth, S. Liu, C. T. Longenecker, R. H. Mackey, K. Matsushita, D. Mozaffarian, M. E. Mussolino, K. Nasir, R. W. Neumar, L. Palaniappan, D. K. Pandey, R. R. Thiagarajan, M. J. Reeves, M. Ritchey, C. J. Rodriguez, G. A. Roth, W. D. Rosamond, C. Sasson, A. Towfighi, C. W. Tsao, M. B. Turner, S. S. Virani, J. H. Voeks, J. Z. Willey, J. T. Wilkins, J. H. Wu, H. M. Alger, S. S. Wong, P. Muntner, C. American Heart Association Statistics, and S. Stroke Statistics. Heart disease and stroke statistics-2017 update: a report from the American Heart Association. Circulation 135:e146–e603, 2017.
Buse, E. E., S. L. Hilbert, R. A. Hopkins, and G. L. Converse. Pulse duplicator hydrodynamic testing of bioengineered biological heart valves. Cardiovasc. Eng. Technol. 7:352–362, 2016.
Converse, G. L., E. E. Buse, K. R. Neill, C. R. McFall, H. N. Lewis, M. C. VeDepo, R. W. Quinn, and R. A. Hopkins. Design and efficacy of a single-use bioreactor for heart valve tissue engineering. J. Biomed. Mater. Res. B Appl. Biomater. 105:249–259, 2017.
Dumont, K., J. Yperman, E. Verbeken, P. Segers, B. Meuris, S. Vandenberghe, W. Flameng, and P. R. Verdonck. Design of a new pulsatile bioreactor for tissue engineered aortic heart valve formation. Artif. Organs 26:710–714, 2002.
Durst, C. A., and K. J. Grande-Allen. Design and physical characterization of a synchronous multivalve aortic valve culture system. Ann. Biomed. Eng. 38:319–325, 2010.
Flanagan, T. C., C. Cornelissen, S. Koch, B. Tschoeke, J. S. Sachweh, T. Schmitz-Rode, and S. Jockenhoevel. The in vitro development of autologous fibrin-based tissue-engineered heart valves through optimised dynamic conditioning. Biomaterials 28:3388–3397, 2007.
Gandaglia, A., A. Bagno, F. Naso, M. Spina, and G. Gerosa. Cells, scaffolds and bioreactors for tissue-engineered heart valves: a journey from basic concepts to contemporary developmental innovations. Eur. J. Cardiothorac. Surg. 39:523–531, 2011.
Ghazanfari, S., A. Driessen-Mol, B. Sanders, P. E. Dijkman, S. P. Hoerstrup, F. P. Baaijens, and C. V. Bouten. In vivo collagen remodeling in the vascular wall of decellularized stented tissue-engineered heart valves. Tissue Eng. Part A 21:2206–2215, 2015.
Hildebrand, D. K., Z. J. Wu, J. E. Mayer, Jr, and M. S. Sacks. Design and hydrodynamic evaluation of a novel pulsatile bioreactor for biologically active heart valves. Ann. Biomed. Eng. 32:1039–1049, 2004.
Hoerstrup, S. P., R. Sodian, J. S. Sperling, J. P. Vacanti, and J. E. Mayer, Jr. New pulsatile bioreactor for in vitro formation of tissue engineered heart valves. Tissue Eng. 6:75–79, 2000.
ISO/TC. 5840-1:2015: cardiovascular implants—cardiac valve prostheses—part 1: general requirements, 2015.
Jana, S., B. J. Tefft, D. B. Spoon, and R. D. Simari. Scaffolds for tissue engineering of cardiac valves. Acta Biomater. 10:2877–2893, 2014.
Jana, S., R. T. Tranquillo, and A. Lerman. Cells for tissue engineering of cardiac valves. J. Tissue Eng. Regen. Med. 10:804–824, 2016.
Kaasi, A., I. A. Cestari, N. A. Stolf, A. A. Leirner, O. Hassager, and I. N. Cestari. A new approach to heart valve tissue engineering: mimicking the heart ventricle with a ventricular assist device in a novel bioreactor. J. Tissue Eng. Regen. Med. 5:292–300, 2011.
Konig, F., T. Hollweck, S. Pfeifer, B. Reichart, E. Wintermantel, C. Hagl, and B. Akra. A pulsatile bioreactor for conditioning of tissue-engineered cardiovascular constructs under endoscopic visualization. J. Funct. Biomater. 3:480–496, 2012.
Lichtenberg, A., I. Tudorache, S. Cebotari, S. Ringes-Lichtenberg, G. Sturz, K. Hoeffler, C. Hurscheler, G. Brandes, A. Hilfiker, and A. Haverich. In vitro re-endothelialization of detergent decellularized heart valves under simulated physiological dynamic conditions. Biomaterials 27:4221–4229, 2006.
Migneco, F., S. J. Hollister, and R. K. Birla. Tissue-engineered heart valve prostheses: ‘state of the heart’. Regen. Med. 3:399–419, 2008.
Mol, A., N. J. Driessen, M. C. Rutten, S. P. Hoerstrup, C. V. Bouten, and F. P. Baaijens. Tissue engineering of human heart valve leaflets: a novel bioreactor for a strain-based conditioning approach. Ann. Biomed. Eng. 33:1778–1788, 2005.
Mol, A., M. C. Rutten, N. J. Driessen, C. V. Bouten, G. Zund, F. P. Baaijens, and S. P. Hoerstrup. Autologous human tissue-engineered heart valves: prospects for systemic application. Circulation 114:I152–158, 2006.
Morsi, Y. S., W. W. Yang, A. Owida, and C. S. Wong. Development of a novel pulsatile bioreactor for tissue culture. J. Artif. Organs 10:109–114, 2007.
Narita, Y., K. Hata, H. Kagami, A. Usui, M. Ueda, and Y. Ueda. Novel pulse duplicating bioreactor system for tissue-engineered vascular construct. Tissue Eng. 10:1224–1233, 2004.
Ramaswamy, S., S. M. Boronyak, T. Le, A. Holmes, F. Sotiropoulos, and M. S. Sacks. A novel bioreactor for mechanobiological studies of engineered heart valve tissue formation under pulmonary arterial physiological flow conditions. J. Biomech. Eng. 136:121009, 2014.
Ramaswamy, S., D. Gottlieb, G. C. Engelmayr, Jr, E. Aikawa, D. E. Schmidt, D. M. Gaitan-Leon, V. L. Sales, J. E. Mayer, Jr, and M. S. Sacks. The role of organ level conditioning on the promotion of engineered heart valve tissue development in-vitro using mesenchymal stem cells. Biomaterials 31:1114–1125, 2010.
Rath, S., M. Salinas, A. G. Villegas, and S. Ramaswamy. Differentiation and distribution of marrow stem cells in flex-flow environments demonstrate support of the valvular phenotype. PLoS ONE 10:e0141802, 2015.
Schenke-Layland, K., F. Opitz, M. Gross, C. Doring, K. J. Halbhuber, F. Schirrmeister, T. Wahlers, and U. A. Stock. Complete dynamic repopulation of decellularized heart valves by application of defined physical signals-an in vitro study. Cardiovasc. Res. 60:497–509, 2003.
Sierad, L. N., A. Simionescu, C. Albers, J. Chen, J. Maivelett, M. E. Tedder, J. Liao, and D. T. Simionescu. Design and testing of a pulsatile conditioning system for dynamic endothelialization of polyphenol-stabilized tissue engineered heart valves. Cardiovasc. Eng. Technol. 1:138–153, 2010.
Spoon, D. B., B. J. Tefft, A. Lerman, and R. D. Simari. Challenges of biological valve development. Interv. Cardiol. 5:319–334, 2013.
VeDepo, M., E. Buse, R. Quinn, R. Hopkins, and G. Converse. Extended bioreactor conditioning of mononuclear cell-seeded heart valve scaffolds. J. Tissue Eng. 9:2041731418767216, 2018.
Warnock, J. N., S. Konduri, Z. He, and A. P. Yoganathan. Design of a sterile organ culture system for the ex vivo study of aortic heart valves. J. Biomech. Eng. 127:857–861, 2005.
Williams, A., S. Nasim, M. Salinas, A. Moshkforoush, N. Tsoukias, and S. Ramaswamy. A “sweet-spot” for fluid-induced oscillations in the conditioning of stem cell-based engineered heart valve tissues. J. Biomech. 65:40–48, 2017.
Ziegelmueller, J. A., E. K. Zaenkert, R. Schams, S. Lackermair, C. Schmitz, B. Reichart, and R. Sodian. Optical monitoring during bioreactor conditioning of tissue-engineered heart valves. ASAIO J. 56:228–231, 2010.
The authors gratefully acknowledge Dr. Sorin V. Pislaru, M.D., Ph.D. for assistance with echocardiographic imaging and Drs. Robert T. Tranquillo and Zeeshan Syedain for assistance with pulse duplicator validation.
Conflict of interest
The authors disclose that Harvard Apparatus owns intellectual property filings related to this work with JB, HH, and JC listed as co-inventors. BT, JC, MY, RH, DM, DD, RS, and AL declare that they have no conflict of interest.
This work was supported by a generous gift from HH Sheikh Hamed Bin Zayed Al-Nahyan and by the NIH (T32 HL007111, K99 HL129068).
This article does not contain any studies with human participants or animals performed by any of the authors.
Associate Editor Ajit P. Yoganathan and Jonathan Butcher oversaw the review of this article.
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Tefft, B.J., Choe, J.A., Young, M.D. et al. Cardiac Valve Bioreactor for Physiological Conditioning and Hydrodynamic Performance Assessment. Cardiovasc Eng Tech 10, 80–94 (2019). https://doi.org/10.1007/s13239-018-00382-2
- Tissue engineered heart valve
- Biomechanical stimulation
- Biochemical stimulation
- Three-dimensional tissue culture
- ISO 5840