去细胞化
细胞外基质
弹性蛋白
化学
十二烷基硫酸钠
质谱成像
多糖
间质细胞
层粘连蛋白
纤维蛋白
组织工程
基质(化学分析)
基底膜
质谱法
色谱法
生物化学
生物医学工程
细胞生物学
蛋白多糖
生物
病理
医学
作者
Giulia Remaggi,Fulvio Barbaro,Giusy Di Conza,Giovanna Trevisi,Carlo Bergonzi,Roberto Toni,Lisa Elviri
出处
期刊:Tissue Engineering Part C-methods
[Mary Ann Liebert]
日期:2022-03-31
卷期号:28 (4): 148-157
被引量:1
标识
DOI:10.1089/ten.tec.2021.0191
摘要
Collagens, elastin, fibrillin, decorin, and laminin are key constituents of the extracellular matrix and basement membrane of mammalian organs. Thus, changes in their quantities may influence the mechanochemical regulation of resident cells. Since maintenance of a native stromal composition is a requirement for three-dimensional (3D) matrix-based recellularization techniques in tissue engineering, we studied the influence of the decellularization detergents on these proteins in porcine kidney, liver, pancreas, and skin. Using a quick thawing/quick microwave-assisted decellularization protocol and two different detergents, sodium dodecyl sulfate (SDS) vs Triton X-100 (TX100), at identical concentration, variations in matrix conservation of stromal proteins were detected by liquid chromatography-mass spectrometry coupled to light and scanning electron microscopies, in dependence on each detergent. In all organs tested except pancreas, collagens were retained to a statistically significant level using the TX100-based protocol. In contrast fibrillin, elastin (except in kidney), and decorin (only in liver) were better preserved with the SDS-dependent protocol. Irrespective of the detergent used, laminin always remained at an irrelevant level. Our results prompt attention to the type of detergent in organ decellularization, suggesting that its choice may influence morphoregulatory inputs peculiar to the type of 3D bioartificial mammalian organ to be reconstructed. Impact statement Simple change of the protocol's main detergent leads to a very substantial difference in the panel of the stromal proteins detected by qualitative and semiquantitative mass spectrometry in acellular porcine matrices. This remarkable methodological variable promises to yield proteomic reference panels in a number of different species-specific acellular matrices allowing for selective retainment of peculiar mechanochemical inputs, to differently address the development of the seeded cells in relation to the type of organ to be bioartificially reconstructed.
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