Department of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Department of Chemistry, CICECO-Aveiro Institute of Materials, University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.
Biomacromolecules. 2020 Dec 14;21(12):4923-4932. doi: 10.1021/acs.biomac.0c01155. Epub 2020 Oct 25.
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.
在人体中，高度组织化的组织依赖于基底膜 (BMs) 的分隔效应，这种分隔效应将不同类型的细胞分隔开来。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)，这两种物质是天然 BMs 的主要成分，可用于三维 (3D) 组织中的细胞分隔。然而，这种分隔结构只能维持 3 天，可能是由于降解问题。在这项研究中，通过转谷氨酰胺酶在 3D 组织中对 Col-IV/LM 层层 (LbL) 纳米膜进行原位交联，制备了一种稳定的 A-BM。通过使用各种流体力学直径的聚苯乙烯纳米颗粒 (PS NPs) 和葡聚糖以及白蛋白，全面研究了分子大小和构象对获得的 A-BM 渗透性的影响。研究结果与已知的肾小球基底膜的尺寸选择性行为非常吻合。交联的 Col-IV/LM 纳米膜表现出更好的稳定性和更强的屏障效应，可维持有组织的 3D 组织的细胞分隔。这种体外 A-BM 在设计更复杂的分隔 3D 组织、理解通过 BMs 进行的独特的细胞-细胞串扰，以及为新药物筛选和其他体外生理研究提供更可靠的 3D 组织模型方面具有巨大的潜力。
