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开发一种猪肾细胞外基质支架作为肾脏再生的平台。

Development of a porcine renal extracellular matrix scaffold as a platform for kidney regeneration.

作者信息

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

机构信息

Department of Urology, School of Medicine, Kyungpook National University, Daegu, Korea.

出版信息

J Biomed Mater Res A. 2015 Apr;103(4):1391-403. doi: 10.1002/jbm.a.35274. Epub 2014 Jul 28.

Abstract

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.

摘要

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。

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