Detergent Decellularization of Heart Valves for Tissue Engineering: Toxicological Effects of Residual Detergents on Human Endothelial Cells

TLDR

Detergents are powerful agents for tissue decellularization, yet residual detergent toxicity can limit their clinical use. This study evaluated how effectively detergents can be removed from decellularized pulmonary valves and the impact of repopulating the resulting scaffolds with human endothelial cells. Porcine pulmonary valves were treated with 1 % sodium deoxycholate, 1 % sodium dodecyl sulfate, or a 0.5 %/0.5 % mixture, washed through ten cycles with detergent concentrations measured by SPE‑HPLC and cytotoxicity assessed by MTS assays, and then reseeded with endothelial cells. All groups achieved efficient decellularization; detergent levels dropped below 50 mg L⁻¹ after 6, 8, and 4 wash cycles for groups A, B, and C, respectively, with no significant cytotoxicity, allowing successful endothelial cell reseeding.

Abstract

Abstract 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.