Type IV collagen conforms to the organization of polylaminin adsorbed on planar substrata
Type IV collagen conforms to the organization of polylaminin adsorbed on planar substrata
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DOI：
10.1016/j.actbio.2020.05.021
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发表时间：
2020-07-15
影响因子：
9.7
通讯作者：
Altankov, George
中科院分区：
文献类型：
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作者：
Coelho-Sampaio, Tatiana;Tenchov, Boris;Altankov, George
组织工程需要开发模拟天然细胞外基质（ECM）的支架。尽管在获得合成间质ECM方面取得了成功，但仍然没有实现人工基底膜（BM）的产生，所述人造基底膜是ECM的专门薄片，其对于大多数组织和内部器官的功能组织是关键的。长期的目标是开发一个平坦的BM样结构，在这里，我们研究了在酸性条件下与层粘连蛋白（LM）同时组装过程中酸溶性Col IV的行为。其基本原理是先前观察到的酸触发LM聚合现象，从而产生可以吸附在基底上的仿生聚层粘连蛋白（polyLM）。出乎意料的是，我们发现Col IV（在酸性条件下不溶胀）容易地结合到polyLM层中，形成在很大程度上模仿在微米尺度下可观察到的单个polyLM的特征多边形形态的网络。扫描量热法和光散射测量支持polyLM和Col IV可以直接相互作用的概念。使用人角质形成细胞（HACAT）和脐静脉内皮细胞（HUVEC）的拟议的人工BM样结构的生物学特性进行了表征。HACAT在混合polyLM/Col IV层上形成分层细胞层，但在Matrigel上不形成分层细胞层，在单独的LM或Col IV上也不形成分层细胞层，而HUVEC改善了皮质F-肌动蛋白和polyLM/Col IV上的紧密连接组织。因此，建议的人工BM再现不仅形态，但也有一些功能特性的天然BM。声明的SignificanceBasement膜（BM）是平坦的生物基质中分离的组织隔间在体内。它们独特的片状结构被认为是由层粘连蛋白和IV型胶原蛋白两个独立的蛋白质网络的结合引起的。在追求人工BM的发展时，我们发现，当与酸诱导的聚合层粘连蛋白混合时，胶原IV立即符合层粘连蛋白的形状。这意味着蛋白质网络可能不像迄今为止所认为的那样独立组装，而是层粘连蛋白可能命令IV型胶原的组装。我们的混合基质在结构上比商业BM提取物Matrigel更稳定，并且与后者不同，支持体外形成接近天然表皮组织的角质形成细胞分层。(C)2020 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Tissue engineering demands the development of scaffolds that mimic natural extracellular matrices (ECM). Despite the success in obtaining synthetic interstitial ECM, the production of an artificial basement membrane (BM), the specialized thin sheet of ECM that is pivotal for the functional organization of most tissues and internal organs, is still not achieved. With the long-term aim of developing a flat BM-like structure here we investigated the behavior of acid-soluble Col IV during simultaneous assembly with laminin (LM) in acidic conditions. The underlying rationale was the previously observed phenomenon of acid-triggered LM polymerization, giving rise to biomimetic polylaminin (polyLM) that can be adsorbed on the substrate. Unexpectedly, we found that Col IV (that does not polymerize in acidic conditions) readily incorporated into the polyLM layer, forming a network that mimics to a great extent the characteristic polygonal morphology of single polyLM observable at micrometric scale. Scanning calorimetry and light scattering measurements supported the notion that polyLM and Col IV could directly interact. The biological properties of the proposed artificial BM-like structure were characterized using human keratinocytes (HACAT) and umbilical vein endothelial cells (HUVEC). HACAT formed stratified cell layers on the hybrid polyLM/Col IV layer, but not on Matrigel, nor on LM or Col IV alone, while HUVEC improved cortical F-actin and tight juctions organization on polyLM/Col IV. Thus, the proposed artificial BM reproduces not only morphological but also some functional properties of the natural BM.Statement of SignificanceBasement membranes (BMs) are flat biological matrices separating tissue compartments in the body. Their peculiar sheet-like structure is thought to result from the association of two independent protein networks of laminin and collagen IV. While pursuing the development of an artificial BM, we found that, when mixed with acid-induced polymerized laminin, collagen IV immediately conformed to the laminin shape. This implies that the protein networks may not be independently assembled as believed so far, but instead that laminin may command the assembly of collagen IV. Our hybrid matrix was structurally more stable than the commercial BM extract Matrigel and, unlike the latter, supported in vitro formation of a stratified layer of keratinocytes that approximated the organization of the natural epidermis. (C) 2020 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.