Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
Comprehensive Transplant Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
J Tissue Eng Regen Med. 2018 Mar;12(3):e1704-e1715. doi: 10.1002/term.2604. Epub 2017 Dec 10.
Detergents such as sodium dodecyl sulfate (SDS) are commonly used to extract cells from tissues in a process called "decellularization". Residual SDS is difficult to completely remove and may lead to an undesirable host response towards an implanted biomaterial. In this study, we developed a modification for SDS cell extraction from muscle equally efficient to previous methods but leading to significantly less residual SDS remnants in the matrices. Muscle-derived matrices were prepared via 2 SDS-based decellularization methods, which led to removal of either 81.4% or 98.4% of the SDS. In vitro, matrices were seeded with thp1 macrophages and primary human foreskin fibroblasts. By Day 2, both matrices demonstrated similar macrophage polarization; however, fibroblasts cultured on matrices with greater residual SDS expressed higher levels of mRNA associated with fibroblast activation: α-smooth muscle actin and connective tissue growth factor. In vivo, Collagen I gels spiked with increasing concentrations of SDS displayed a corresponding decrease in cell infiltration when implanted subcutaneously in rats after 4 days. Finally, as a model for muscle regeneration, matrices produced by each method were implanted in rat latissimus dorsi defects. At POD 30 greater levels of IL-1β mRNA were present in defects treated with matrices containing higher levels of SDS, indicating a more severe inflammatory response. Although matrices containing higher levels of residual SDS became encapsulated by POD 30 and showed evidence of a foreign body response, matrices with the lower levels of SDS integrated into the defect area with lower levels of inflammatory and fibrosis-related gene expression.
清洁剂，如十二烷基硫酸钠（SDS），常用于从组织中提取细胞，这个过程称为“脱细胞化”。残留的 SDS 很难完全去除，可能会导致植入生物材料后宿主产生不良反应。在这项研究中，我们开发了一种 SDS 细胞提取的改良方法，与之前的方法同样有效，但在基质中残留的 SDS 残余物明显减少。肌肉衍生的基质是通过 2 种基于 SDS 的脱细胞化方法制备的，这两种方法分别去除了 81.4%或 98.4%的 SDS。在体外，基质中接种了 thp1 巨噬细胞和原代人包皮成纤维细胞。在第 2 天，两种基质均表现出类似的巨噬细胞极化；然而，在残留 SDS 水平较高的基质上培养的成纤维细胞表达了更高水平的与成纤维细胞活化相关的 mRNA：α-平滑肌肌动蛋白和结缔组织生长因子。在体内，随着 SDS 浓度的增加，胶原 I 凝胶植入大鼠皮下 4 天后，细胞浸润相应减少。最后，作为肌肉再生的模型，用每种方法制备的基质植入大鼠背阔肌缺陷处。在 POD 30 时，用含有更高 SDS 水平的基质处理的缺陷中存在更高水平的 IL-1β mRNA，表明炎症反应更严重。尽管含有较高残留 SDS 的基质在 POD 30 时被包裹，并显示出异物反应的迹象，但 SDS 水平较低的基质与较低水平的炎症和纤维化相关基因表达一起整合到缺陷区域。
