A new approach to heart valve tissue engineering

 

Willem Flameng

Cardiac Surgery, University Clinic Gasthuisberg Leuven, Belgium.

 

Tissue engineering is aiming to create viable tissue or organ grafts that do self propagate and repair (Grikscheit and Vacanti, 2002) . Three strategies to achieve this goal have been proposed: implantation of cells, in situ tissue regeneration and implantation of in vitro assembled constructs from cells and scaffolds (Griffith and Naughton, 2002) . In the field of cardiac valve tissue engineering the last mentioned strategy has been generally opted for (Bader et al., 1998;Grikscheit and Vacanti, 2002;Terada et al., 2000) .

Considering the matrix materials again two directions have been followed since the first paper by Shinoka and coworkers (1995) on tissue engineering of a replacement heart valve. A first choice was the construction of heart valves from biodegradable materials. Here again, Shinoka (1995) and some other groups (Sodian et al., 2000;Hoerstrup et al., 1998) focused on seeding of scaffolds comprised of artificial biodegradable material such as polyglycolic acid meshes and polyhydroxyalkanoate, a bacterium-derived thermoplastic polyester. Whereas the first scaffold was highly porous, too stiff to create a complete valve (Shinoka et al., 1995) and potentially immunogenic, the second was more flexible but required special salt leaching preparations to obtain an acceptable porosity. Different approaches either using no scaffold (Ye et al., 2000b) or human fibrin gel based scaffold (Ye et al., 2000a) have also been examined since they would overcome the specific problems of the artificial scaffold materials. Yet another branch of cardiac valve tissue engineering is using acellularised human (Dohmen et al., 2002a;Dohmen et al., 2002b) or porcine aortic root (Bader et al., 1998;Dohmen et al., 2002b;Dohmen et al., 2001) as a matrix material. This material has the great advantage that it is a functional valve in itself and does not has to be manufactured. Some groups have been seeding these matrices in vitro (Bader et al., 1998) , whereas others have mainly focused on the acellularisation process, claiming that these matrices show spontaneous repopulation in vivo (O'Brien et al., 1999) .

Although all these matrices showed promising results, serious drawbacks have also been reported. Considering the artificial scaffolds rapid hydrolysation can result in construct breakdown (Cebotari et al., 2002;Shinoka et al., 1998) whereas implantation of xenogeneic acellular aorta in animals resulted in aneurysmal dilatation (Allaire et al., 1994) . Also recent clinical trials using implants of non-seeded but acellularised porcine valves showed catastrophic results (Simon et al., 2002) related to early degeneration of these matrices.

In vitro seeding has major methodological problems besides those of cell seeding itself (eg. sterility, cell culturing, etc.). The loss of seeded cells is one of the major problems when in vitro seeding is used (Jansson et al., 2001;Rademacher et al., 2001) and to overcome these problems different types of flow/pressure systems have been designed. They range from stretching devices or simple flow systems (Ziegler and Nerem, 1994) to fully functional bioreactors gradually imitating the real haemodynamic flow and pressure stresses (Hoerstrup et al., 2000;Sodian et al., 2002;Dumont et al., 2002) .

At present our group has addressed another approach, originating from a first attempt to grow an artificial artery graft out of granulation tissue (Simmons et al., 1975) . This graft, the Sparks mandril prosthesis, was constructed from the tissue encapsulating a subcutaneously implanted plastic/glass rod. Although the cellular component was removed and the remaining tissue tanned, this is to our knowledge the first attempt to construct a vascular prosthesis from granulation tissue. More recently Campbell and coworkers (1999) showed the construction of a functional vascular graft from a capsule generated from an intraperitoneally deposited capsule around an implanted silastic tube. The everted the capsule to internalise the exterior mesothelial layer and implanted the grafts in the abdominal aorta of both rats and rabbits. The type of capsule that is deposited, is comprised out of an internal layer of macophages, some layers of (myo)fibroblasts and an external layer of mesothelial cells and is generally described as ‘Foreign Body Reaction’ (Butler et al., 2001a;Butler et al., 2001b) . At present it has already been proven that the entire layer of (myo)fibroblasts is of haematopoietic origin (Campbell et al., 2000) , as was shown by reconstituting the bone marrow of female irradiated mice with male derived transplanted cells. In a later stage of their research the same group (Bayes-Genis et al., 2002) claimed that the myofibroblasts were derived from the macrophages, but also reported the presence of an undifferentiated cell population in the explants. This was done by studying the cell morphology and colocalisation of ASMA and CD172a. Although ASMA is a generally used marker for myofibroblast (Della Rocca et al., 2000;Taylor et al., 2000;DeRuiter et al., 1997) , the CD172a is not an exclusive marker for macrophages but is also expressed in a subpopulation of haematopoietic stem cells (Kuci et al., 2003;Vogel et al., 2003) . Considering these findings we feel that at this stage we can not exclude the possibility of the presence of stem cells in the granulation tissue. Therefore we chose to pursue this approach further and study the use of “immature” foreign body reaction as a means to repopulate a biological but stabilised matrix. This approach avoids but does not exclude the use of a bioreactor since the seeding is done by the valve recipient itself.

Therefore our group has been working on an alternative matrix comprised of a cross-linked scaffold and an autologous neomatrix covering the foreign scaffold. As a cross-linked scaffold we have used a non cytotoxic photooxidated bovine pericardium (Moore, 2001) , which is shown to tolerate in vitro cell seeding (Jansson et al., 2001) .

 

 

References

1. Allaire E, Guettier C, Bruneval P, Plissonnier D, Michel JB (1994) Cell-free arterial grafts: morphologic characteristics of aortic isografts, allografts, and xenografts in rats. J Vasc Surg 19: 446-456.

2. Bader A, Schilling T, Teebken OE, Brandes G, Herden T, Steinhoff G, Haverich A (1998) Tissue engineering of heart valves--human endothelial cell seeding of detergent acellularized porcine valves. Eur J Cardiothorac Surg 14: 279-284.

3. Bayes-Genis A, Campbell JH, Carlson PJ, Holmes DR, Jr., Schwartz RS (2002) Macrophages, myofibroblasts and neointimal hyperplasia after coronary artery injury and repair. Atherosclerosis 163: 89-98.

4. Butler K, Benghuzzi H, Tucci S (2001a) Tissue-implant response following soft tissue implantation of poly-L-lysine coated UHMW-polyethylene into adult male rats. Biomed Sci Instrum 37: 19-24.

5. Butler KR, Benghuzzi HA, Puckett A (2001b) Morphometric evaluation of tissue-implant reaction associated with ALCAP and TCP bioceramics in vivo. J Invest Surg 14: 139-152.

6. Campbell JH, Efendy JL, Campbell GR (1999) Novel vascular graft grown within recipient's own peritoneal cavity. Circ Res 85: 1173-1178.

7. Campbell JH, Efendy JL, Han C, Girjes AA, Campbell GR (2000) Haemopoietic origin of myofibroblasts formed in the peritoneal cavity in response to a foreign body. J Vasc Res 37: 364-371.

8. Cebotari S, Mertsching H, Kallenbach K, Kostin S, Repin O, Batrinac A, Kleczka C, Ciubotaru A, Haverich A (2002) Construction of autologous human heart valves based on an acellular allograft matrix. Circulation 106: I63-I68.

9. Della Rocca F, Sartore S, Guidolin D, Bertiplaglia B, Gerosa G, Casarotto D, Pauletto P (2000) Cell composition of the human pulmonary valve: a comparative study with the aortic valve--the VESALIO Project. Vitalitate Exornatum Succedaneum Aorticum labore Ingegnoso Obtinebitur. Ann Thorac Surg 70: 1594-1600.

10. DeRuiter MC, Poelmann RE, VanMunsteren JC, Mironov V, Markwald RR, Gittenberger-de Groot AC (1997) Embryonic endothelial cells transdifferentiate into mesenchymal cells expressing smooth muscle actins in vivo and in vitro. Circ Res 80: 444-451.

11. Dohmen PM, Lembcke A, Hotz H, Kivelitz D, Konertz WF (2002a) Ross operation with a tissue-engineered heart valve. Ann Thorac Surg 74: 1438-1442.

12. Dohmen PM, Ozaki S, Verbeken E, Yperman J, Flameng W, Konertz WF (2002b) Tissue engineering of an auto-xenograft pulmonary heart valve. Asian Cardiovasc Thorac Ann 10: 25-30.

13. Dohmen PM, Ozaki S, Yperman J, Flameng W, Konertz W (2001) Lack of calcification of tissue engineered heart valves in juvenile sheep. Semin Thorac Cardiovasc Surg 13: 93-98.

14. Dumont K, Yperman J, Verbeken E, Segers P, Meuris B, Vandenberghe S, Flameng W, Verdonck PR (2002) Design of a new pulsatile bioreactor for tissue engineered aortic heart valve formation. Artif Organs 26: 710-714.

15. Griffith LG, Naughton G (2002) Tissue engineering--current challenges and expanding opportunities. Science 295: 1009-1014.

16. Grikscheit TC, Vacanti JP (2002) The history and current status of tissue engineering: The future of pediatric surgery. J Pediatr Surg 37: 277-288.

17. Hoerstrup SP, Sodian R, Sperling JS, Vacanti JP, Mayer JE, Jr. (2000) New pulsatile bioreactor for in vitro formation of tissue engineered heart valves. Tissue Eng 6: 75-79.

18. Hoerstrup SP, Zund G, Schoeberlein A, Ye Q, Vogt PR, Turina MI (1998) Fluorescence activated cell sorting: a reliable method in tissue engineering of a bioprosthetic heart valve. Ann Thorac Surg 66: 1653-1657.

19. Jansson K, Bengtsson L, Swedenborg J, Haegerstrand A (2001) In vitro endothelialization of bioprosthetic heart valves provides a cell monolayer with proliferative capacities and resistance to pulsatile flow. J Thorac Cardiovasc Surg 121: 108-115.

20. Kuci S, Wessels JT, Buhring HJ, Schilbach K, Schumm M, Seitz G, Loffler J, Bader P, Schlegel PG, Niethammer D, Handgretinger R (2003) Identification of a novel class of human adherent CD34- stem cells that give rise to SCID-repopulating cells. Blood 101: 869-876.

21. Moore MA (2001) PhotoFix: unraveling the mystery. J Long Term Eff Med Implants 11: 185-197.

22. O'Brien MF, Goldstein S, Walsh S, Black KS, Elkins R, Clarke D (1999) The SynerGraft valve: a new acellular (nonglutaraldehyde-fixed) tissue heart valve for autologous recellularization first experimental studies before clinical implantation. Semin Thorac Cardiovasc Surg 11: 194-200.

23. Rademacher A, Paulitschke M, Meyer R, Hetzer R (2001) Endothelialization of PTFE vascular grafts under flow induces significant cell changes. Int J Artif Organs 24: 235-242.

24. Shinoka T, Breuer CK, Tanel RE, Zund G, Miura T, Ma PX, Langer R, Vacanti JP, Mayer JE, Jr. (1995) Tissue engineering heart valves: valve leaflet replacement study in a lamb model. Ann Thorac Surg 60: S513-S516.

25. Shinoka T, Shum-Tim D, Ma PX, Tanel RE, Isogai N, Langer R, Vacanti JP, Mayer JE, Jr. (1998) Creation of viable pulmonary artery autografts through tissue engineering. J Thorac Cardiovasc Surg 115: 536-545.

26. Simmons P, Fogarty TJ, Pennell JP (1975) Nineteen months clinical experience with the Sparks mandril graft. J Am Assoc Nephrol Nurses Tech 2: 120-123.

27. Simon, P., Kasimir, M. T., Seebacher, G., Weigel, G., Ullrich, R., Salzer-Muhar, U., Rieder, E., and Wolner, E. Early failure of the tissue engineering porcine heart valve synergraft in pediatric patients. The european association for cardio-thoracic surgery, 16th annual meeting abstractbook.  282. 2002.

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28. Sodian R, Lemke T, Fritsche C, Hoerstrup SP, Fu P, Potapov EV, Hausmann H, Hetzer R (2002) Tissue-engineering bioreactors: a new combined cell-seeding and perfusion system for vascular tissue engineering. Tissue Eng 8: 863-870.

29. Sodian R, Sperling JS, Martin DP, Egozy A, Stock U, Mayer JE, Jr., Vacanti JP (2000) Fabrication of a trileaflet heart valve scaffold from a polyhydroxyalkanoate biopolyester for use in tissue engineering. Tissue Eng 6: 183-188.

30. Taylor PM, Allen SP, Yacoub MH (2000) Phenotypic and functional characterization of interstitial cells from human heart valves, pericardium and skin. J Heart Valve Dis 9: 150-158.

31. Terada S, Sato M, Sevy A, Vacanti JP (2000) Tissue engineering in the twenty-first century. Yonsei Med J 41: 685-691.

32. Vogel W, Grunebach F, Messam CA, Kanz L, Brugger W, Buhring HJ (2003) Heterogeneity among human bone marrow-derived mesenchymal stem cells and neural progenitor cells. Haematologica 88: 126-133.

33. Ye Q, Zund G, Benedikt P, Jockenhoevel S, Hoerstrup SP, Sakyama S, Hubbell JA, Turina M (2000a) Fibrin gel as a three dimensional matrix in cardiovascular tissue engineering. Eur J Cardiothorac Surg 17: 587-591.

34. Ye Q, Zund G, Jockenhoevel S, Hoerstrup SP, Schoeberlein A, Grunenfelder J, Turina M (2000b) Tissue engineering in cardiovascular surgery: new approach to develop completely human autologous tissue. Eur J Cardiothorac Surg 17: 449-454.

35. Ziegler T, Nerem RM (1994) Tissue engineering a blood vessel: regulation of vascular biology by mechanical stresses. J Cell Biochem 56: 204-209.

 

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