Detergent decellularization of heart valves for tissue engineering: toxicological effects of residual detergents on human endothelial cells.
Detergents are powerful agents for tissue decellularization. Despite this, the high toxicity of detergent residua can be a major limitation. This study evaluated the efficacy of detergent removal from decellularized pulmonary valves (PVs) and the consequences of repopulation with human endothelial cells (HECs). Porcine PVs were treated with 1% sodium deoxycholate (SDC), group A; 1% sodium dodecyl sulfate (SDS), group B; and a mixture of 0.5% SDC/0.5% SDS, group C (n = 5 each). After each of 10 succeeding wash cycles (WCs), samples of the washing solution (WS) were analyzed by solid phase extraction and high performance liquid chromatography for the presence of detergents. Metabolic activity of HEC was also assessed in the WS samples (cytotoxicity and MTS assays). Decellularized and washed PVs were reseeded with HEC. Histological analysis demonstrated efficient tissue decellularization in all groups. Detergents' concentration in all WSs decreased exponentially and was below 50 mg/L after 6, 8, and 4 WCs in groups A, B, and C, respectively. This concentration resulted in no significant toxic influence on cell cultures, and scaffolds could be efficiently reseeded with HEC. In conclusion, intensive washing of detergent decellularized valvular scaffolds lowers the residual contamination below a hazardous threshold and allows their successful repopulation with HEC for tissue engineering purposes.
Cebotari
Serghei
S
Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany. cebotari.serghei@mh-hannover.de
Tudorache
Igor
I
Jaekel
Thomas
T
Hilfiker
Andres
A
Dorfman
Suzanne
S
Ternes
Waldemar
W
Haverich
Axel
A
Lichtenberg
Artur
A
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003840:Deoxycholic Acid / Q000633:toxicity*; D003902:Detergents / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D011664:Pulmonary Valve / Q000187:drug effects* / Q000473:pathology; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity*; D052616:Solid Phase Extraction; D013552:Swine; D023822:Tissue Engineering; D054457:Tissue Scaffolds
D003902:Detergents; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate