Precision Medicine Key Laboratory, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
J Biomed Mater Res A. 2021 Sep;109(9):1623-1632. doi: 10.1002/jbm.a.37158. Epub 2021 Mar 7.
Whole-organ engineering is emerging as an alternative source for xenotransplantation in end-stage diseases. Utilization of decellularized whole lung scaffolds created by detergent perfusion is an effective strategy for organ replacement. In the current study, we attempted to decellularize porcine whole lungs to generate an optimal and reproducible decellularized matrix for future clinical use. Porcine whole lungs were decellularized via perfusion of various detergents (sodium dodecyl sulfate (SDS)/Triton X-100, sodium lauryl ether sulfate (SLES)/Triton X-100, dextrose/SDS/Triton X-100 and dextrose/SLES/Triton X-100) through the pulmonary artery and bronchus of the lung. The decellularized scaffolds were evaluated for decellularization efficiency, extracellular matrix (ECM) component preservation, xenoantigen removal and compatibility. The resulting lung scaffolds obtained from treatment with the dextrose/SLES/Triton X-100 cocktail showed minimal residual cellular components and xenoantigens, including DNA and protein, and good preservation of ECM composition. Evaluation of the porcine lung ECM by specific staining and immunofluorescence confirmed that the three-dimensional ultrastructure of the ECM was noticeably preserved in the SLES-treated groups. In addition, the decellularized lung scaffolds originating from the dextrose/SLES/Triton X-100 cocktail supported cell adhesion and growth. In summary, the novel detergent SLES alleviated the damage to retain a better-preserved ECM than SDS. Sequential Triton X-100 perfusion eliminated SLES. Moreover, performing dextrose perfusion in advance further protected scaffold components, especially collagen. We developed an optimal dextrose/SLES/Triton X-100 cocktail method that can be used for the decellularization of porcine whole lung to obtain a clinical-scale bioengineered scaffold.
整体器官工程作为异种移植治疗终末期疾病的替代来源正在兴起。利用去污剂灌注制备的去细胞化全肺支架进行器官替代是一种有效的策略。在本研究中，我们尝试通过肺动静脉和支气管对猪全肺进行去细胞化，以生成最佳且可重复的去细胞化基质，用于未来的临床应用。我们通过肺动静脉和支气管对猪全肺进行去细胞化，使用不同的去污剂（十二烷基硫酸钠（SDS）/Triton X-100、月桂醚硫酸钠（SLES）/Triton X-100、葡萄糖/SDS/Triton X-100 和葡萄糖/SLES/Triton X-100）进行灌注。通过对去细胞化支架的去细胞化效率、细胞外基质（ECM）成分保存、异种抗原去除和相容性进行评估。结果表明，用葡萄糖/SLES/Triton X-100 鸡尾酒处理得到的肺支架细胞残留成分和异种抗原（包括 DNA 和蛋白质）最少，ECM 成分保存良好。通过对猪肺 ECM 的特殊染色和免疫荧光评估证实，SLES 处理组 ECM 的三维超微结构得到了很好的保留。此外，来源于葡萄糖/SLES/Triton X-100 鸡尾酒的去细胞化肺支架支持细胞黏附和生长。总之，新型去污剂 SLES 减轻了损伤，保留了更好的 ECM。连续用 Triton X-100 灌注可消除 SLES。此外，预先进行葡萄糖灌注可进一步保护支架成分，特别是胶原蛋白。我们开发了一种优化的葡萄糖/SLES/Triton X-100 鸡尾酒方法，可用于猪全肺的去细胞化，以获得临床规模的生物工程支架。
