• Journal of Chest Surgery
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• v.37 no.4
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• pp.297-306
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• 2004

Current artificial heart valves have several disadvantages, such as thromboembolism, limited durability, infection, and inability to grow. The solution to these problems would be to develop a tissue-engineered heart valves containing autologous cells. The aim of this study was to optimize the protocol to obtain a porcine acellular matrix and seed goat autologous endothelial cells on it, and to evaluate the biological responses of xenograft and xeno-autograft heart valves in goats. Material and Method: Fresh porcine pulmonic valves were treated with one method among 3 representative decellularization protocols (Triton-X, freeze-thawing, and NaCl-SDS). Goat venous endothelial cells were isolated and seeded onto the acellularized xenograft leaflets. Microscopic examinations were done to select the most effective method of decellularizing xenogeneic cells and seeding autologous endothelial cells. Two pulmonic valve leaflets of. 6 goats were replaced by acellularized porcine leaflets with or without seeding autologous endothelial cells while on cardiopulmonary bypass. Goats were sacrificed electively at 6 hours, 1 day, 1 week, 1 month, 3 months, and 6. months after operation. Morphologic examinations were done to see the biological responses of replaced valve leaflets. Result: The microscopic examinations showed that porcine cells were almost completely removed in the leaflets treated with NaCl-SDS. The seeded endothelial cells were more evenly preserved in NaCl-SDS treatment. All 6 goats survived the operation without complications. The xeno- autografts and xenografts showed the appearance, the remodeling process, and the cellular functions of myofibroblasts, 1 day, 1 month, and 3 months after operation, respectively. They were compatible with the native pulmonary leaflet (control group) except for the increased cellularity at 6 months. The xenografts revealed the new endothelial cell lining at that time. Conclusion: Treatment with NaCl-SDS was most effective in obtaining decellularized xenografts and facilitate seeding autologous endothelial cells. The xenografts and xeno-autografts were repopulated with myofibroblasts and endothelial cells in situ serially. Both of grafts served as a matrix for a tissue engineered heart valve and developed into autologous tissue for 6 months.

• Polymer(Korea)
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• v.29 no.3
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• pp.294-299
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• 2005

Thermo-responsive poly(N-isopropylacrylamide) (PIPAAm) was covalently patterned by masked el electron beam irradiation. Introduction of PIPAAm on tissue culture polystyrene dish was confirmed by ATR-FTIR and ESCA measurements. Hepatocytes were cultured at $37^{circ}C$ on these surfaces. Cells adhered on PIPAAm-grafted domains were detached by reducing culture temperature to $20^{circ}C$. Endothelial cells were then seeded and cultured on the same surfaces. Seeded endothelial cells were selectively attached on hepatocytes detached and PIPAAm-grafted domains and could be co-cultured with hepatocytes on the same culture dishes with clear pattern. This co-culture method enabled long-term co-culture of hepatocytes with endothelial cells.

• Journal of Chest Surgery
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• v.37 no.6
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• pp.482-491
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• 2004

Background: The xenogenic or allogenic valves after in Vitro repopulation with autologous cells or in vivo repo-pulation after acellularization treatment to remove the antigenicity could used as an alternative to synthetic polymer scaffold. In the present study, we evaluated the process of repopulation by recipient cell to the acellu-larized xenograft treated with NaCl-SDS solution and grafted in the right ventricular outflow tract. Material and Method: Porcine pulmonary valved conduit were treated with. NaCl-SDS solution to make the grafts acellularized and implanted in the right ventricular outflow tract of the goats under cardiopulmonary bypass. After evaluating the functions of pulmonary valves by echocardiography, goats were sacrificed at 1 week, 1 month, 3 months, 6 months, and 12 months after implantation, respectively. After retrieving the implanted valved conduits, histopathologic examination with Hematoxylin-Eosin, Masson' trichrome staining and immunohistochemical staining was performed. Result: Among the six goats, which had been implanted with acellularized pulmonary valved conduits, five survived the expected time period. Echocardiographic examinations for pulmonary valves revealed good function except mild regurgitation and stenosis. Microscopic analysis of the leaflets showed progressive cellular in-growth, composed of fibroblasts, myofibroblasts, and endothelial cells, into the acellularized leaflets over time. Severe inflammatory respon-se was detected in early phase, though it gradually decreased afterwards. The extracellular matrices were regenerated by repopulated cells on the recellularized portion of the acellularized leaflet. Conclusion: The acellularized xenogenic pulmonary valved conuits were repopulated with fibroblasts, myofibroblasts, and endothelial cells of the recipient and extracellullar matrices were regenerated by repopulted cells 12 months after the implantation. The functional integrity of pulmonary valves was well preserved. This study showed that the acellularized porcine xenogenic valved conduits could be used as an ideal valve prosthesis with long term durability.