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J Thorac Cardiovasc Surg 2004;127:399-405
© 2004 The American Association for Thoracic Surgery

Surgery for acquired cardiovascular disease

Decellularization protocols of porcine heart valves differ importantly in efficiency of cell removal and susceptibility of the matrix to recellularization with human vascular cells

^a Department of Cardiothoracic Surgery and Ludwig-Boltzmann-Institute for Cardiosurgical Research, University of Vienna, Vienna, Austria

Received for publication March 22, 2003; revisions received May 1, 2003; accepted for publication June 23, 2003.

^* Address for reprints: Guenter Weigel, MD, Department of Cardiothoracic Surgery, University of Vienna, AKH, Waehringer Guertel 18-20, 1090 Vienna, Austria
guenter.weigel{at}akh-wien.ac.at

OBJECTIVE: We compared 3 different decellularization protocols in porcine heart valves for efficiency of complete cell removal and potential for recellularization.

METHODS: Porcine aortic and pulmonary roots were treated with trypsin, sodium-dodecyl-sulphate, or a new method using 0.25% tert-octylphenyl-polyoxyethylen in combination with sodium-deoxycholate. After a subsequent ribonuclease digestion, specimens were seeded with in vitro expanded human saphenous vein endothelial cells and myofibroblasts.

RESULTS: After treatment with trypsin and subsequent ribonuclease digestion, endothelial attachment took place; however, xenogenic cells were still visible within the matrix. Unexpectedly, when human cells were seeded onto specimens that had been decellularized with sodium-dodecyl-sulphate, the matrices were surrounded by nonviable endothelial cell fragments, indicating a toxic influence of the ionic detergent; 0.25% tert-octylphenyl-polyoxyethylen together with sodium-deoxycholate completely removed porcine cells and enabled host recellularization.

CONCLUSION: Compared with trypsin and sodium-dodecyl-sulphate involving decellularization procedures, reported to be effective in cell removal and susceptible to recellularization with human cells, only the porcine matrix treated with a new detergent-based decellularization method using 0.25% tert-octylphenyl-polyoxyethylen/sodium-deoxycholate followed by nuclease digestion presented an excellent scaffold for recellularization with human cells.

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