[en] Pulmonary valve replacement is performed with excellent resultant hemodynamics in patients that have underlying congenital or acquired heart valve defects. Despite recent advancements in right ventricular outflow tract reconstruction, an increased risk of developing infective endocarditis remains, which has a more common occurrence for conduits of bovine jugular vein (BJV) origin compared with cryopreserved homografts. The reason for this is unclear although it is hypothesized to be associated with an aberrant phenotypic state of cells that reendothelialize the graft tissue postimplantation. The aim of this study was to develop an in vitro model that enables the analysis of endothelial cell (EC) attachment to cardiac graft tissues under flow. In the experiments, EC attachment was optimized on bovine pericardium (BP) patch using human umbilical vein ECs. Different biological coatings, namely gelatin, fibronectin, plasma, or a combination of fibronectin and plasma were tested. After cell adaptation, graft tissues were exposed to laminar flow in a parallel-plate flow chamber. Cell retention to the tissue was analyzed after nuclear staining with YO-PRO-1 and a membranous localization of VE-cadherin. Experiments showed that combined coating with fibronectin and blood plasma together with a two-phased shear pattern resulted in a relevant cell monolayer on BP patch and cryopreserved homograft. For BJV tissue, no adherent cells under both static and shear conditions were initially observed. In conclusion, having established the new flow chamber system we could obtain EC layers on the surface of BP patch and cryopreserved pulmonary homograft tissues. The presented in vitro system can serve as a competent model to study cell phenotypes on cardiac grafts in the close-to-physiologic environment. Moreover, this approach allows broad applications and enables further development by testing more complex conditions.
Moreillon, P., and Que, Y.A. Infective endocarditis. Lancet 363, 139, 2004.
Toyoda, N., Chikwe, J., Itagaki, S., Gelijns, A.C., Adams, D.H., and Egorova, N.N. Trends in infective endocarditis in California and New York state, 1998-2013. J Am Med Assoc 317, 1652, 2017.
Van Dijck, I., Budts, W., Cools, B., et al. Infective endocarditis of a transcatheter pulmonary valve in comparison with surgical implants. Heart 101, 788, 2015.
Zilla, P., Fullard, L., Trescony, P., et al. Glutaraldehyde detoxification of aortic wall tissue: a promising perspective for emerging bioprosthetic valve concepts. J Heart Valve Dis 6, 510, 1997.
Bengtsson, L., Radegran, K., and Haegerstrand, A. In vitro endothelialization of commercially available heart valve bioprostheses with cultured adult human cells. Eur J Cardio-Thoracic Surg 7, 393, 1993.
Jayakrishnan, A., and Jameela, S.R. Glutaraldehyde as a fixative in bioprostheses and drug delivery matrices. Biomaterials 17, 471, 1996.
Jansson, K., Bengtsson, L., Swedenborg, J., and Haegerstrand, A. In vitro endothelialization of bioprosthetic heart valves provides a cell monolayer with proliferative capacities and resistance to pulsatile flow. J Thorac Cardiovasc Surg 121, 108, 2001.
Eybl, E., Griesmacher, A., Grimm, M., and Wolner, E. Toxic effects of aldehydes released from fixed pericardium on bovine aortic endothelial cells. J Biomed Mater Res 23, 1355, 1989.
Umashankar, P.R., Mohanan, P.V., and Kumari, T.V. Glutaraldehyde treatment elicits toxic response compared to decellularization in bovine pericardium. Toxicol Int 19, 51, 2012.
Eberl, T., Siedler, S., Schumacher, B., Zilla, P., Schlaudraff, K., and Fasol, R. Experimental in vitro endothelialization of cardiac valve leaflets. Ann Thorac Surg 53, 487, 1992.
Bin, F., Yinglong, L., Nin, X., Kai, F., Laifeng, S., and Xiaodong, Z. Construction of tissue-engineered homograft bioprosthetic heart valves in vitro. ASAIO J 52, 303, 2006.
Zavazava, N., Simon, A., Sievers, H.H., Bernhard, A., and Müller-Ruchholtz, W. Porcine valves are reendothelialized by human recipient endothelium in vivo. J Thorac Cardiovasc Surg 109, 702, 1995.
Vincentelli, A., Latrémouille, C., Zegdi, R., et al. Does glutaraldehyde induce calcification of bioprosthetic tissues? Ann Thorac Surg 66, S255, 1998.
Grabenwöger, M., Grimm, M., Eybl, E., et al. Decreased tissue reaction to bioprosthetic heart valve material after L-glutamic acid treatment. A morphological study. J Biomed Mater Res 26, 1231, 1992.
McCarron, J.G., Lee, M.D., and Wilson, C. The endothelium solves problems that endothelial cells do not know exist. Trends Pharmacol Sci 38, 322, 2017.
Becker, B.F., Heindl, B., Kupatt, C., and Zahler, S. Endothelial function and hemostasis. Z Kardiol 89, 160, 2000.
Van Hinsbergh, V.W.M. Endothelium-role in regulation of coagulation and inflammation. Semin Immunopathol 34, 93, 2012.
Trantina-Yates, A.E., Human, P., Bracher, M., and Zilla, P. Mitigation of bioprosthetic heart valve degeneration through biocompatibility: in vitro versus spontaneous endothelialization. Biomaterials 22, 1837, 2001.
Sigler, M. Endothelialisation of cardiovascular implants? A matter of concern. J Clin Exp Cardiolog 04, e130, 2013.
Lopez-Moya, M., Melgar-Lesmes, P., Kolandaivelu, K., De La Torre Hernández, J.M., Edelman, E.R., and Balcells, M. Optimizing glutaraldehyde-fixed tissue heart valves with chondroitin sulfate hydrogel for endothelialization and shielding against deterioration. Biomacromol Am Chem Soc 19, 1234, 2018.
Guldner, N.W., Jasmund, I., Zimmermann, H., et al. Detoxification and endothelialization of glutaraldehyde-fixed bovine pericardium with titanium coating a new technology for cardiovascular tissue engineering. Circulation 119, 1653, 2009.
Jansson, K., Bengtsson, L., Swedenborg, J., and Haegerstrand, A. In vitro endothelialization of bioprosthetic heart valves provides a cell monolayer with proliferative capacities and resistance to pulsatile flow. J Thorac Cardiovasc Surg 121, 108, 2001.
Hutcheson, J.D., Blaser, M.C., and Aikawa, E. Giving calcification its due: recognition of a diverse disease: a first attempt to standardize the field. Circ Res 120, 270, 2017.
Liesenborghs, L., Meyers, S., Lox, M., et al. Staphylococcus aureus endocarditis: distinct mechanisms of bacterial adhesion to damaged and inflamed heart valves. Eur Heart J 40, 3248, 2019.
Sánchez, P.F., Brey, E.M., and Bricenõ, J.C. Endothelialization mechanisms in vascular grafts. J Tissue Eng Regen Med 12, 2164, 2018.
Vogel, M., Franke, J., Frank, W., and Schroten, H. Flow in the well: computational fluid dynamics is essential in flow chamber construction. Cytotechnology 55, 41, 2007.
Ditkowski, B., Leeten, K., Jashari, R., Jones, E., and Heying, R. Staphylococcus aureus adheres avidly to decellularised cardiac homograft tissue in vitro in the fibrinogen-dependent manner. Cardiol Young 30, 1783, 2020.
Ditkowski, B., Veloso, T.R., Bezulska-Ditkowska, M., et al. An In Vitro model of a parallel-plate perfusion system to study bacterial adherence to graft tissues. J Vis Exp 2019, e58476, 2019.
Ditkowski, B., Bezulska-Ditkowska, M., Jashari, R., et al. Antiplatelet therapy abrogates platelet-assisted Staphylococcus aureus infectivity of biological heart valve conduits. J Thorac Cardiovasc Surg 19, 33112, 2019.
Kasimir, M.T., Weigel, G., Sharma, J., et al. The decellularized porcine heart valve matrix in tissue engineering: platelet adhesion and activation. Thromb Haemost 94, 562, 2005.
Esch, M.B., Post, D.J., Shuler, M.L., and Stokol, T. Characterization of in vitro endothelial linings grown within microfluidic channels. Tissue Eng A 17, 2965, 2011.
Teebken, O.E., Bader, A., Steinhoff, G., and Haverich, A. Tissue engineering of vascular grafts: human cell seeding of decellularised porcine matrix. Eur J Vasc Endovasc Surg 19, 381, 2000.
Salacinski, H.J., Tai, N.R., Punshon, G., Giudiceandrea, A., Hamilton, G., and Seifalian, A.M. Optimal endothelialisation of a new compliant poly(carbonate-urea)urethane vascular graft with effect of physiological shear stress. Eur J Vasc Endovasc Surg 20, 342, 2000.
Kent, K.C., Oshima, A., Ikemoto, T., and Whittemore, A.D. An in vitro model for human endothelial cell seeding of a small diameter vascular graft. ASAIO Trans 34, 578, 1988.
Row, S., Santandreu, A., Swartz, D.D., and Andreadis, S.T. Cell-free vascular grafts: recent developments and clinical potential. Technology 05, 13, 2017.
Cardinal, K.O.H., Bonnema, G.T., Hofer, H., Barton, J.K., and Williams, S.K. Tissue-engineered vascular grafts as in vitro blood vessel mimics for the evaluation of endothelialization of intravascular devices. Tissue Eng 12, 3431, 2006.
Liu, H., Gong, X., Jing, X., et al. Shear stress with appropriate time-step and amplification enhances endothelial cell retention on vascular grafts. J Tissue Eng Regen Med 11, 2965, 2017.
Uzarski, J.S., Van De Walle, A.B., and Mcfetridge, P.S. In vitro method for real-time, direct observation of cellvascular graft interactions under simulated blood flow. Tissue Eng C Methods 20, 116, 2014.
Melchiorri, A.J., Bracaglia, L.G., Kimerer, L.K., Hibino, N., and Fisher, J.P. In Vitro Endothelialization of Biode-gradable Vascular Grafts Via Endothelial Progenitor Cell Seeding and Maturation in a Tubular Perfusion System Bioreactor. Tissue Eng C Methods 22, 663, 2016.
Burke, F.M., McCormack, N., Rindi, S., Speziale, P., and Foster, T.J. Fibronectin-binding protein B variation in Staphylococcus aureus. BMC Microbiol 10, 160, 2010.
Peel, M.M., and DiMilla, P.A. Effect of Cell-Cell Interactions on the Observable Strength of Adhesion of Sheets of Cells. Ann Biomed Eng 27, 236, 1999.
Kesler, K.A., Herring, M.B., Arnold, M.P., et al. Enhanced strength of endothelial attachment on polyester elastomer and polytetrafluoroethylene graft surfaces with fibronectin substrate. J Vasc Surg 3, 58, 1986.
Vohra, R., Thomson, G.J.L., Carr, H.M.H., Sharma, H., and Walker, M.G. Comparison of different vascular prostheses and matrices in relation to endothelial seeding. Br J Surg 78, 417, 1991.
Dejana, E., Lampugnani, M., Giorgi, M., Gaboli, M., and Marchisio, P. Fibrinogen induces endothelial cell adhesion and spreading via the release of endogenous matrix proteins and the recruitment of more than one integrin receptor. Blood Am Soc Hematol 75, 1509, 1990.
Recek, N., Mozetic, M., Jaganjac, M., Milkovic, L., Zarkovic, N., and Vesel, A. Adsorption of proteins and cell adhesion to plasma treated polymer substrates. Int J Polym Mater Polym Biomater 63, 685, 2014.
Xiao, Y., and Isaacs, S.N. Enzyme-linked immunosorbent assay (ELISA) and blocking with bovine serum albumin (BSA)-not all BSAs are alike. J Immunol Methods 384, 148, 2012.
Harris, E.S., and Nelson, W.J. VE-cadherin: at the front, center, and sides of endothelial cell organization and function. Curr Opin Cell Biol 22, 651, 2010.
Jashari, R., Van Hoeck, B., Tabaku, M., and Vanderkelen, A. Banking of the human heart valves and the arteries at the European homograft bank (EHB)-overview of a 14-year activity in this International Association in Brussels. Cell Tissue Bank 5, 239, 2004.
Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., and Walter, P. Blood vessels and endothelial cells. In: Dries, D.J., ed. Molecular Biology of the Cell. New York: Garland Science, 2002.
Neethling, W.M.L., Puls, K., and Rea, A. Comparison of physical and biological properties of CardioCel with commonly used bioscaffolds. Interact Cardiovasc Thorac Surg 26, 985, 2018.
Medtronic Integrated Valved Conduit for RVOT [Internet]. Available at: https://www.accessdata.fda.gov/cdrh-docs/pdf2/H020003C.pdf (accessed January 15, 2021).
Synovis Supple Peri-Guard [Internet]. Available at: https://advancedsurgery.baxter.com/sites/g/files/ebysai1521/files/2019-01/SUPPLE-PERI-GUARD-IFU.pdf (accessed January 15, 2021).
