Pediatric Urology and Regenerative Medicine Research Center, Children's Medical Center, Pediatric Center of Excellence, Tehran University of Medical Sciences, Tehran, Iran.
Tracheal Diseases Research Center (TDRC), National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Tissue Cell. 2020 Oct;66:101396. doi: 10.1016/j.tice.2020.101396. Epub 2020 Jun 2.
The main goal of this study was to assess the effect of different sterilization treatment for sterilization of decellularized kidney tissue. Rabbit kidneys were decellularized by the perfusion-based method using sodium dodecyl sulfate (SDS) and Triton X-100. Then, decellularized kidney slices were prepared and sterilized by an antibiotic cocktail, PAA (0.5 %, 1% and 1.5 %), 5KG γ-irradiation and 320-480 nm UV-irradiation. Histological evaluations, DNA quantification assay, MTT assay, scanning electron microscopy (SEM), mechanical test and bacterial and fungal culture tests were performed to determine the quality of decellularization and sterilization processes. The kidney slices were seeded by adipose-derived mesenchymal stem cells (ASCs) to assess the cell adhesion capability after treatment. The results of the current study indicated that PAA 0.5 % was the most efficient method to completely decontaminate rabbit decellularized kidney tissue while preserving the mechanical properties and main components of the matrix which are necessary for cell-matrix interaction and cell adhesion. The 5KG γ-irradiation was determined to be the most destructive sterilization method, with reduced the mechanical strengths as well as altered microstructure of the kidney matrix and no cell adhesion. In addition, UV-irradiation is not able to sterile the decellularized tissues. Therefore PAA 0.5 % sterilization method can be a powerful means for sterilization of biological scaffolds.
本研究的主要目的是评估不同灭菌处理方法对脱细胞肾脏组织的灭菌效果。采用基于灌注的方法，使用十二烷基硫酸钠（SDS）和 Triton X-100 对兔肾脏进行脱细胞处理。然后，制备脱细胞肾切片，并通过抗生素混合物、PAA（0.5%、1%和 1.5%）、5KG γ 射线和 320-480nm UV 射线进行灭菌。进行组织学评估、DNA 定量测定、MTT 测定、扫描电子显微镜（SEM）、力学试验和细菌及真菌培养试验，以确定脱细胞和灭菌过程的质量。将脂肪来源间充质干细胞（ASCs）接种到肾切片上，以评估处理后细胞黏附能力。本研究结果表明，PAA 0.5%是最有效的方法，可以完全清除兔脱细胞肾脏组织的污染物，同时保留基质的机械性能和主要成分，这是细胞-基质相互作用和细胞黏附所必需的。5KG γ 射线被确定为最具破坏性的灭菌方法，会降低肾脏基质的机械强度和改变其微观结构，且没有细胞黏附。此外，UV 射线不能使脱细胞组织无菌。因此，PAA 0.5%的灭菌方法可能是生物支架灭菌的有效手段。
