Choi Seock Hwan, Chun So Young, Chae Seon Yeong, Kim Jin Rae, Oh Se Heang, Chung Sung Kwang, Lee Jin Ho, Song Phil Hyun, Choi Gyu-Seog, Kim Tae-Hwan, Kwon Tae Gyun
Department of Urology, School of Medicine, Kyungpook National University, Daegu, Korea.
J Biomed Mater Res A. 2015 Apr;103(4):1391-403. doi: 10.1002/jbm.a.35274. Epub 2014 Jul 28.
Acellular scaffolds, possessing an intact three-dimensional extracellular matrix (ECM) architecture and biochemical components, are promising for regeneration of complex organs, such as the kidney. We have successfully developed a porcine renal acellular scaffold and analyzed its physical/biochemical characteristics, biocompatibility, and kidney reconstructive potential. Segmented porcine kidney cortexes were treated with either 1% (v/v) Triton X-100 (Triton) or sodium dodecyl sulfate (SDS). Scanning electron microscopy showed both treatments preserved native tissue architecture, including porosity and composition. Swelling behavior was higher in the Triton-treated compared with the SDS-treated scaffold. Maximum compressive strength was lower in the Triton-treated compared with the SDS-treated scaffold. Attenuated total reflective-infrared spectroscopy showed the presence of amide II (-NH) in both scaffolds. Furthermore, richer ECM protein and growth factor contents were observed in the Triton-treated compared with SDS-treated scaffold. Primary human kidney cell adherence, viability, and proliferation were enhanced on the Triton-treated scaffold compared with SDS-treated scaffold. Following murine in vivo implantation, tumorigenecity was absent for both scaffolds after 8 weeks and in the Triton-treated scaffold only, glomeruli-like structure formation and neovascularity were observed. We identified 1% Triton X-100 as a more suitable decellularizing agent for porcine renal ECM scaffolds prior to kidney regeneration.
脱细胞支架具有完整的三维细胞外基质(ECM)结构和生化成分,在诸如肾脏等复杂器官的再生方面很有前景。我们已成功开发出一种猪肾脱细胞支架,并分析了其物理/生化特性、生物相容性和肾脏重建潜力。将分段的猪肾皮质用1%(v/v)的 Triton X-100(Triton)或十二烷基硫酸钠(SDS)处理。扫描电子显微镜显示两种处理均保留了天然组织结构,包括孔隙率和组成。与SDS处理的支架相比,Triton处理的支架的肿胀行为更高。与SDS处理的支架相比,Triton处理的支架的最大抗压强度更低。衰减全反射红外光谱显示两种支架中均存在酰胺II(-NH)。此外,与SDS处理的支架相比,在Triton处理的支架中观察到更丰富的ECM蛋白和生长因子含量。与SDS处理的支架相比,原代人肾细胞在Triton处理的支架上的黏附、活力和增殖增强。在小鼠体内植入后,8周后两种支架均无致瘤性,仅在Triton处理的支架中观察到肾小球样结构形成和新生血管。我们确定1% Triton X-100是肾脏再生前猪肾ECM支架更合适的去细胞剂。