Detergent Decellularization of Heart Valves for Tissue Engineering: Toxicological Effects of Residual Detergents on Human Endothelial Cells
Detergent Decellularization of Heart Valves for Tissue Engineering: Toxicological Effects of Residual Detergents on Human Endothelial Cells
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DOI：
10.1111/j.1525-1594.2009.00796.x
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发表时间：
2010-03-01
影响因子：
2.4
通讯作者：
Lichtenberg, Artur
中科院分区：
文献类型：
--
作者：
Cebotari, Serghei;Tudorache, Igor;Lichtenberg, Artur
洗涤剂是组织脱细胞的强力剂。尽管如此，洗涤剂残留物的高毒性可能是一个主要限制。本研究评估了去除率去除脱细胞肺动脉瓣（pv）的效果和人内皮细胞（HECs）再生的后果。A组用1%脱氧胆酸钠（SDC）处理猪pv；1%十二烷基硫酸钠（SDS）， B组；0.5% SDC/0.5% SDS的混合物为C组（n = 5）。在10个洗涤循环（WCs）之后，通过固相萃取和高效液相色谱分析洗涤溶液（WS）的样品，以确定洗涤剂的存在。在WS样品中也评估了HEC的代谢活性（细胞毒性和MTS测定）。去细胞和洗净的pv用HEC重新播种。组织学分析显示，所有组均有有效的组织脱细胞。A、B、C组洗涤剂浓度分别在6、8、4次洗涤后呈指数下降，均低于50 mg/L。该浓度对细胞培养没有明显的毒性影响，并且支架可以有效地重新播种HEC。总之，对去污剂脱细胞瓣膜支架进行密集洗涤可将残留污染降低到危险阈值以下，并允许其成功地与HEC一起重新填充，用于组织工程目的。
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.