Department of Basic Sciences, Histology Section, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
Tissue Engineering Lab, Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
J Biomed Mater Res A. 2019 Nov;107(11):2576-2588. doi: 10.1002/jbm.a.36763. Epub 2019 Aug 9.
In situ recellularization of the liver decellularized scaffold is a potential therapeutic alternative for liver transplantation. We aimed to develop an in situ procedure for recellularization of the rat liver using sodium lauryl ether sulfate (SLES) compared with Triton X-100/SDS. Rat liver specimens were rinsed with PBS, decellularized with either Triton X-100/SDS or SLES, and finally rinsed by distilled water. The efficiency of decellularized liver scaffolds was evaluated by histological, confocal Raman microscopy, histochemical staining, and DNA quantification assessments. Finally, in vivo studies were done to assess the biocompatibility of the liver scaffold by serum biochemical parameters and the recellularization capacity by histological and immunohistochemistry staining. Findings confirmed the preservation of extracellular matrix (ECM) components such as reticular, collagen, glycosaminoglycans, and neutral carbohydrates in both Triton X-100/SDS- and SLES-treated livers. Hoechst, feulgen, Hematoxylin and eosin, and DNA quantification assessments confirmed complete genetic content removal. The serological parameters showed no adverse impact on the liver functions. Transplantation of SLES-treated cell-free decellularized liver showed extensive neovascularization along with migration of the fibrocytes and adipocytes and some immune cells. Also, immunohistochemical staining showed that the oval cells, stellate cells, cholangiocytes and hepatocytes invaded extensively into the graft. It is concluded that SLES can be considered as a promising alternative in the liver decellularization process, and the transplanted decellularized liver can appropriately be revascularized and regenerated.
肝去细胞支架的原位再细胞化是肝移植的一种有潜力的治疗选择。我们旨在开发一种使用十二烷基醚硫酸钠（SLES）原位再细胞化大鼠肝脏的方法，与 Triton X-100/SDS 进行比较。大鼠肝标本用 PBS 冲洗，用 Triton X-100/SDS 或 SLES 去细胞化，最后用蒸馏水冲洗。通过组织学、共聚焦拉曼显微镜、组织化学染色和 DNA 定量评估来评估去细胞化肝支架的效率。最后，通过血清生化参数和组织学及免疫组织化学染色评估肝支架的生物相容性进行体内研究。研究结果证实，Triton X-100/SDS 和 SLES 处理的肝脏中保留了细胞外基质（ECM）成分，如网状、胶原、糖胺聚糖和中性碳水化合物。Hoechst、Feulgen、苏木精和伊红以及 DNA 定量评估证实了完全去除遗传物质。血清学参数表明对肝功能没有不良影响。SLES 处理的无细胞去细胞化肝移植后，随着纤维细胞和脂肪细胞以及一些免疫细胞的迁移，出现广泛的新生血管形成。免疫组织化学染色显示，卵圆细胞、星状细胞、胆管细胞和肝细胞广泛侵入移植物。总之，SLES 可以被认为是肝去细胞化过程中的一种有前途的替代方法，移植的去细胞化肝可以适当进行再血管化和再生。
