Serghei Cebotari1, I. Tudorache2, Thomas Jaekel3, Andres Hilfiker2, Suzanne E. Dorfman2, Waldemar Ternes3, Axel Haverich2, Artur Lichtenberg2
1Department of Cardiothoracic Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany
2Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, and
3Department of Chemical Analytics, Centre for Food Science, School of Veterinary Medicine, Hannover, Germany
Tóm tắt
AbstractDetergents 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.
