In Vitro Modelling of Tissue using Isolated Vascular Cells on a Synthetic Collagen Matrix as a Substitute for Heart Valves

Background: Tissue engineering is a promising approach for obtaining lifetime durability in biological heart valves. Basic questions with respect to the selection of suitable cell populations as well as scaffolds remain unsolved. The purpose of this study was to develop a tissue-like substitute in vitro for replacement of diseased valves in vivo. Methods: Smooth-muscle cells (SMCs) were isolated from human and porcine aortic tissue using the ‘explant technique’ and endothelial cells from collagenase digestion. Seeding and cultivation of isolated cells was performed on a type-I collagen matrix. The scaffold-cell specimen was investigated using light and electron microscopy. Cupromeronic blue and immunoprecipitation were used for ultracytochemical staining. Results: SMCs were allowed to grow to multilayers and migrate into the collagen network. We found a tissue-like morphology in these samples characterised by several layers of cells, spaces between the cell layers filled with newly formed extracellular matrix components, compartmentalisation of proteoglycans and their association with fibrilar matrix and the cell surface. Endothelium cells covered the SMCs of the scaffold with a histological topography similar to heart valves. Conclusions: This is an approach for in vitro modelling of tissue-like substitutes and preparing plane multicellular tissues as substitutes for heart valves. This model may also be used for cell biological investigations of cell-matrix interactions.

Tissue engineering - Heart valve - Collagen scaffold - Glycosaminoglycans - Extracellular matrix - Proteoglycans

References

• 1 Carabello B A, Crawford F A. Valvular heart disease.  N Engl J Med. 1997;  337 32-41
• 2 Vongpatanasin W, Hillis D, Lange R A. Prosthetic heart valves.  N Engl J Med. 1996;  335 407-416
• 3 Jamieson W R. Modern cardiac valve devices -bioprostheses and mechanical prostheses: State of the art.  J Cardiac Surg. 1993;  8 89-98
• 4 Cannegieter S C, Rosendaal F R, Briet E. Thromboembolic and bleeding complications in patients with mechanical heart valve prostheses.  Circ. 1994;  89 635-641
• 5 Sodian R, Hoerstrup S P, Sperling J S, Daebritz S H, Martin D P, Schoen F J, Vacanti J P, Mayer J E. Tissue engineering of heart valves: in vitro experiences.  Ann Thorac Surg. 2000;  70 140-144
• 6 Sodian R, Hoerstrup S P, Sperling J S, Martin D P, Daebritz S, Mayer J E, Vacanti J P. Evaluation of biodegradable, three-dimensional matrices for tissue engineering of heart valves.  ASAIO Journal. 2000;  46 107-110
• 7 Shinoka T, Breuer C, Tanel R E, Zund G, Miura T, Ma P X, Langer R, Vacanti J P, Mayer J E. Tissue engineering heart valves: valve leaflet replacement study in a lamb model.  Ann Thorac Surg. 1995;  60 S 513-516
• 8 Taylor P M, Allen S P, Yacoub M H. Phenotypic and functional characterization of interstitial cells from human heart valves, pericardium and skin.  J of Heart Valve Disease. 2000;  9 150-158
• 9 Schoof H, Bruns L, Apel J, Heschel I, Rau G. Einfluß des Einfriervorganges auf die Porenstruktur gefriergetrockneter Kollagenschwämme.  DKV-Berichte. 1997;  24 (Band III) 40-57
• 10 Vischer P, Buddecke E. Alteration of glycosyltransferase activities during proliferation of cultivated arterial endothelial cells and smooth muscle cells.  Exp Cell Res. 1985;  158 15-28
• 11 Schön P, Vischer P, Völker W, Schmidt A, Faber V. Cell-associated proteoheparan sulfate mediates binding and uptake of thrombospondin in cultured porcine vascular endothelial cells.  Eur J Cell Biol. 1992;  59 329-339
• 12 Völker W. Cytochemical characterization and mapping of glycoconjugates in arterial tissues and vascular cell cultures by light and electron microscopy. In: H. Robenek, NJ. Servers (eds.) Cell Interactions In Artherosclerosis. CRC Press, Boca Raton 1992: 181-206
• 13 Völker W, Schmidt A, Buddecke E. Compartmentation and characterization of different proteoglycans in bovine arterial wall.  J Histochem Cytochem. 1986;  34 1293-1299
• 14 Völker W, Schmidt A, Buddecke E. Mapping of proteoglycans in human arterial tissue.  Eur J Cell Biol. 1987;  45 72-79
• 15 Vischer P, Buddecke E. Different action of heparin and fucoidan on arterial smooth muscle cell proliferation and thrombospondin and fibronectin metabolism.  Eur J Cell Biol. 1991;  56 407-414
• 16 Hammermeister K E, Sethi G K, Henderson W G, Oprian C, Kim T, Rahimtoola S. A comparison of outcomes in men 11 years after heart-valve replacement with a mechanical valve or bioprosthesis.  N Engl J Med. 1993;  18 1289-1296
• 17 Shoen F J, Levy R J. Tissue heart valves: current challenges and future research perspectives. Founder's Award, Twenty-fifth Annual Meeting of the Society for Biomaterials, Providence, RJ. 1999; April 28 - May 2
• 18 McGiffin D C, Galbraith A J, O'Brien M F, McLachlan G J, Naftel D C, Adams P, Reddy S, Early L. An analysis of valve re-replacement after aortic valve replacement with biologic devices.  J Thorac Cardiovasc Surg. 1997;  113 311-318
• 19 Schoen F J, Levy R J, Piehler H R. Pathological considerations in replacement cardiac valves.  Cardiovasc Pathol. 1992;  1 29-52
• 20 Miller D C. Predictors of outcome in patients with prosthetic valve endocarditis (PVE) and potential advantages of homograft aortic root replacement for prosthetic ascending aortic valve-graft infections.  J Cardiac Surg. 1990;  5 53-62
• 21 Rajani B, Mee R B, Ratliff N B. Evidence for rejection of homografts cardiac valves in infants.  J Thorac Cardiovasc Surg. 1998;  115 111-117
• 22 Shum-Tim D, Stock U, Hrkach J, Shinoka T, Lien J, Moses M A, Stamp A, Taylor G, Moran A M, Landis W, Langer R, Vacanti J P, Mayer J E. Tissue engineering of autologous aorta using a new biodegradable polymer.  Ann Thorac Surg. 1999;  68 2298-2305
• 23 Mayer J E, Shinoka T, Shum-Tim D. Tissue engineering of cardiovascular structures.  Current Opinion in Cardiology. 1997;  12 528-532
• 24 Zund G, Ye S, Hoerstrup S P, Schoeberlein A, Schmid A C, Grunenfelder J, Vogt P, Turina M. Tissue engineering in cardiovascular surgery: MTT, a rapid and reliable quantitative method to assess the optimal human cell seeding on polymeric meshes.  Eur J Cardiothorac Surg. 1999;  15 519-524
• 25 Ye Q, Zund G, Jockenhoevel S, Hoerstrup S P, Schoeberlein A, Grunenfelder J, Turina M. Tissue engineering in cardiovascular surgery: new approach to develop completely human autologous tissue.  Eur J Cardiothorac Surg. 2000;  17 449-454
• 26 Ishihora T, Ferrans V J, Jones M, Boyce S W, Kawanami O, Robert W C. Histologic and ultrastructural features of normal human parietal pericardium.  Am J Cardiol. 1980;  46 744-753
• 27 Juliano R L, Haskill S. Signal transduction from the extracellular matrix.  J Cell Biol. 1993;  120 577-585