In Situ Cross-Linking of Artificial Basement Membranes in 3D Tissues and Their Size-Dependent Molecular Permeability
In Situ Cross-Linking of Artificial Basement Membranes in 3D Tissues and Their Size-Dependent Molecular Permeability
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3D 组织中人工基底膜的原位交联及其尺寸依赖性分子渗透性
DOI：
10.1021/acs.biomac.0c01155
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发表时间：
2020
影响因子：
6.2
通讯作者：
Matsusaki Michiya
中科院分区：
文献类型：
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作者：
Zeng Jinfeng;Correia Clara R.;Mano Jo?o F.;Matsusaki Michiya
在人体中，高度组织化的组织依赖于基底膜（BM）的区室化效应，该效应将不同类型的细胞分开。我们最近报道了一种由IV型胶原和层粘连蛋白（Col-IV/LM）组成的人工基底膜（A-BM），它们是天然BM的主要成分，用于三维（3D）组织中的细胞区室化。然而，这种分隔结构只能维持3天，可能是由于退化问题。在这项研究中，通过转氨酶在3D组织中原位交联Col-IV/LM逐层（LbL）纳米膜来制造稳健的A-BM。利用聚苯乙烯纳米颗粒（PS NPs）和具有各种流体动力学直径的葡聚糖以及白蛋白，对分子大小和构型对所获得的A-BM的渗透性的影响进行了全面的研究。这些发现与已知的肾小球基底膜的尺寸选择性行为一致。交联的Col-IV/LM纳米膜表现出改善的稳定性和更强大的屏障作用，以维持有序3D组织的细胞区室化。这种体外A-BM在设计更复杂的区室化3D组织、理解通过BM的独特细胞-细胞串扰以及为新药筛选和其他体外生理学研究提供更可靠的3D组织模型方面表现出巨大的潜力。
In the human body, highly organized tissues rely on the compartmentalization effect of basement membranes (BMs) that separate different types of cells. We recently reported an artificial basement membrane (A-BM) composed of type-IV collagen and laminin (Col-IV/LM), which are the main components of natural BMs, for cell compartmentalization in three-dimensional (3D) tissues. However, such compartmentalized structures can be maintained only for 3 days, probably due to the degradation issues. In this study, a robust A-BM was fabricated by in situ cross-linking the Col-IV/LM layer-by-layer (LbL) nanofilms in 3D tissues by transglutaminase. The effects of molecular size and configuration on the permeability of obtained A-BMs were comprehensively studied using polystyrene nanoparticles (PS NPs) and dextran with various hydrodynamic diameters, as well as albumin. The findings agreed well with the known size-selective behavior of the glomerular basement membrane. Cross-linked Col-IV/LM nanofilms demonstrate improved stability and a more powerful barrier effect to maintain cell compartmentalization for organized 3D tissues. This in vitro A-BM exhibit great potentials for the design of more complex compartmentalized 3D tissues, for understanding the unique cell–cell cross talk through BMs, and for providing a more reliable 3D tissue model for new drug screening and other in vitro physiological studies.