使用低浓度十二烷基硫酸钠对猪颈动脉进行脱细胞处理。
Decellularization of porcine carotid arteries using low-concentration sodium dodecyl sulfate.
机构信息
Department of Vascular Surgery, Xuanwu Hospital, Capital Medical University, Beijing, P.R. China.
出版信息
Int J Artif Organs. 2021 Jul;44(7):497-508. doi: 10.1177/0391398820975420. Epub 2020 Nov 23.
BACKGROUND
The decellularized scaffold is a promising material for producing tissue-engineered vascular grafts (TEVGs) because of its complex, native-like three-dimensional structure and mechanical properties. Sodium dodecyl sulfate (SDS), one of the most commonly used decellularization reagents, appears to be more effective than other detergents for removing cells from dense tissues. The concentrations of SDS used in previous studies and their effects on decellularization are not consistent.
METHODS
In this study, porcine carotid arteries were decellularized using detergent-based protocols using Triton X-100 followed by SDS at different concentrations and exposing time. Cell removal efficiency and composition were evaluated by histological analysis, and DNA and collagen quantification. Ultrastructure, mechanical properties, pore size distribution, and in vivo biocompatibility of decellularized arteries were also evaluated.
RESULTS
The DNA content of decellularized scaffolds treated with 0.3% SDS for 72 h or 0.5% SDS for 48 h was significantly less than that treated with 1% SDS for 30 h. There was a significant loss of soluble collagen after treatment with 1% SDS relative to native arteries. The extensive loss of elastin and glycosaminoglycans was observed in decellularized arteries treated with 0.5% SDS or 1% SDS. The basement membrane and biomechanics were also damaged by these two protocols. Moreover, decellularized scaffolds became more porous with many large pores after treatment with 0.3% SDS.
CONCLUSION
Low-concentration SDS could be a suitable choice for artery decellularization. Decellularized porcine carotid arteries, prepared using Triton X-100 followed by 0.3% SDS, may be a promising biological scaffold for TEVGs.
背景
脱细胞支架因其复杂的、类似天然的三维结构和机械性能，是制造组织工程血管移植物（TEVGs）的有前途的材料。十二烷基硫酸钠（SDS）是最常用的脱细胞试剂之一，似乎比其他去污剂更有效地从致密组织中去除细胞。以前研究中使用的 SDS 浓度及其对脱细胞的影响并不一致。
方法
在这项研究中，使用基于去污剂的方案，使用 Triton X-100 后用不同浓度和暴露时间的 SDS 对猪颈动脉进行脱细胞处理。通过组织学分析、DNA 和胶原蛋白定量评估细胞去除效率和组成。还评估了脱细胞动脉的超微结构、力学性能、孔径分布和体内生物相容性。
结果
用 0.3% SDS 处理 72 小时或 0.5% SDS 处理 48 小时的脱细胞支架的 DNA 含量明显低于用 1% SDS 处理 30 小时的含量。用 1% SDS 处理后，可溶性胶原蛋白明显丢失。用 0.5% SDS 或 1% SDS 处理的脱细胞动脉中观察到弹性蛋白和糖胺聚糖大量丢失。基底膜和生物力学也被这两种方案破坏。此外，用 0.3% SDS 处理后，脱细胞支架变得更加多孔，出现许多大孔。
结论
低浓度 SDS 可能是动脉脱细胞的合适选择。用 Triton X-100 预处理后用 0.3% SDS 处理的脱细胞猪颈动脉可能是 TEVGs 的有前途的生物支架。
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