Influence of pH on Extracellular Matrix Preservation During Lung Decellularization
Influence of pH on Extracellular Matrix Preservation During Lung Decellularization
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DOI：
10.1089/ten.tec.2013.0492
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发表时间：
2014-12-01
影响因子：
3
通讯作者：
Niklason, Laura E.
中科院分区：
文献类型：
--
作者：
Tsuchiya, Tomoshi;Balestrini, Jenna L.;Niklason, Laura E.
用于再生医学的脱细胞器官的产生需要保存器官细胞外基质（ECM），作为为接种到器官支架上的细胞的分化和迁移提供关键线索的手段。本研究的目的是评估在大鼠肺去细胞化过程中不同pH水平对关键ECM组分保存的影响。在本文中，我们表明，pH值的3-[（3-胆酰胺丙基）二甲基铵]-1-丙磺酸（CHAPS）为基础的脱细胞溶液影响ECM保留，细胞去除，也是潜在的宿主反应后植入无细胞肺组织。ECM组分的保存，包括弹性蛋白、纤连蛋白和层粘连蛋白，在测试的较低pH条件下（测试的pH范围：8、10、12）得到更好的保留;糖胺聚糖在较低pH组中也得到更高程度的保存。大鼠肺脱细胞后的DNA含量与脱细胞溶液的pH呈负相关。尽管细胞骨架蛋白和显着的残留DNA的检测水平，在pH 8脱细胞的组织表现出最大的组织结构的维护和最少的诱导所有无细胞条件下的主机响应。这些结果突出了pH对通过器官脱细胞化获得的结果的影响，并且表明基于关键ECM组分的保存和植入结果，改变用于脱细胞化的溶液的pH可能影响细胞适当分化和归巢到支架内的适当位置的能力。
The creation of decellularized organs for use in regenerative medicine requires the preservation of the organ extracellular matrix (ECM) as a means to provide critical cues for differentiation and migration of cells that are seeded onto the organ scaffold. The purpose of this study was to assess the influence of varying pH levels on the preservation of key ECM components during the decellularization of rat lungs. Herein, we show that the pH of the 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS)-based decellularization solution influences ECM retention, cell removal, and also the potential for host response upon implantation of acellular lung tissue. The preservation of ECM components, including elastin, fibronectin, and laminin, were better retained in the lower pH conditions that were tested (pH ranges tested: 8, 10, 12); glycosaminoglycans were preserved to a higher extent in the lower pH groups as well. The DNA content following decellularization of the rat lung was inversely correlated with the pH of the decellularization solution. Despite detectible levels of cyotoskeletal proteins and significant residual DNA, tissues decellularized at pH 8 demonstrated the greatest tissue architecture maintenance and the least induction of host response of all acellular conditions. These results highlight the effect of pH on the results obtained by organ decellularization and suggest that altering the pH of the solutions used for decellularization may influence the ability of cells to properly differentiate and home to appropriate locations within the scaffold, based on the preservation of key ECM components and implantation results.