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


CONCLUSIONS
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.
Rieder
Erwin
E
Department of Cardiothoracic Surgery, University of Vienna, Austria.
Kasimir
Marie-Theres
MT
Silberhumer
Gerd
G
Seebacher
Gernot
G
Wolner
Ernst
E
Simon
Paul
P
Weigel
Guenter
G
D001021:Aortic Valve / Q000166:cytology* / Q000601:surgery* / Q000648:ultrastructure; D001672:Biocompatible Materials / Q000494:pharmacology; D001705:Bioprosthesis; D002614:Chelating Agents / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000494:pharmacology; D003851:Deoxyribonucleases / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000737:chemistry* / Q000187:drug effects / Q000648:ultrastructure; D006350:Heart Valve Prosthesis; D006801:Humans; D008854:Microscopy, Electron; D017830:Octoxynol / Q000494:pharmacology; D011474:Prosthesis Design; D011664:Pulmonary Valve / Q000166:cytology* / Q000601:surgery* / Q000648:ultrastructure; D012260:Ribonucleases / Q000494:pharmacology; D012501:Saphenous Vein / Q000166:cytology / Q000648:ultrastructure; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology
D001672:Biocompatible Materials; D002614:Chelating Agents; D003902:Detergents; D013501:Surface-Active Agents; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol; D004492:Edetic Acid; D003851:Deoxyribonucleases; D012260:Ribonucleases