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筛选胶原蛋白 IV 片段自组装成稳定结构，可能对再生医学有用。

Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine.

机构信息

General Command of the Polish Armed Forces, Medical Division, Zwirki i Wigury 103/105, 00-912 Warsaw, Poland.

Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, Narutowicza 60, 90-136 Lodz, Poland.

出版信息

Int J Mol Sci. 2021 Dec 18;22(24):13584. doi: 10.3390/ijms222413584.

DOI:10.3390/ijms222413584
PMID:34948383
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8708666/
Abstract

The aim of the research was to check whether it is possible to use fragments of type IV collagen to obtain, as a result of self-assembling, stable spatial structures that could be used to prepare new materials useful in regenerative medicine. Collagen IV fragments were obtained by using DMT/NMM/TosO as a coupling reagent. The ability to self-organize and form stable spatial structures was tested by the CD method and microscopic techniques. Biological studies covered: resazurin assay (cytotoxicity assessment) on BJ, BJ-5TA and C2C12 cell lines; an alkaline version of the comet assay (genotoxicity), Biolegend Legendplex human inflammation panel 1 assay (SC cell lines, assessment of the inflammation activity) and MTT test to determine the cytotoxicity of the porous materials based on collagen IV fragments. It was found that out of the pool of 37 fragments (peptides - and -) reconstructing the outer sphere of collagen IV, nine fragments (peptides: , , , , , , , and ), as a result of self-assembling, form structures mimicking the structure of the triple helix of native collagens. The stability of spatial structures formed as a result of self-organization at temperatures of 4 °C, 20 °C, and 40 °C was found. The application of the MST method allowed us to determine the K of binding of selected fragments of collagen IV to ITGα1β1. The stability of the spatial structures of selected peptides made it possible to obtain porous materials based on their equimolar mixture. The formation of the porous materials was found for cross-linked structures and the material stabilized only by weak interactions. All tested peptides are non-cytotoxic against all tested cell lines. Selected peptides also showed no genotoxicity and no induction of immune system responses. Research on the use of porous materials based on fragments of type IV collagen, able to form stable spatial structures as scaffolds useful in regenerative medicine, will be continued.

摘要

本研究旨在探讨是否可以使用 IV 型胶原片段，通过自组装获得稳定的空间结构，从而制备可用于再生医学的新材料。我们使用 DMT/NMM/TosO 作为偶联试剂获得 IV 型胶原片段。通过 CD 方法和显微镜技术测试其自组织和形成稳定空间结构的能力。生物学研究包括：在 BJ、BJ-5TA 和 C2C12 细胞系上进行 resazurin 测定（细胞毒性评估）；碱性彗星试验（遗传毒性）、Biolegend Legendplex 人类炎症面板 1 测定（SC 细胞系，评估炎症活性）和 MTT 试验，以确定基于 IV 型胶原片段的多孔材料的细胞毒性。结果发现，在所研究的 37 个片段（肽-和-）中，有 9 个片段（肽：、、、、、、、和）通过自组装形成结构，模拟天然胶原三螺旋的结构。在 4°C、20°C 和 40°C 的温度下，发现自组织形成的空间结构具有稳定性。应用 MST 方法，我们能够确定所选 IV 型胶原片段与 ITGα1β1 的结合 Kd。所选肽的空间结构稳定性使得能够获得基于其等摩尔混合物的多孔材料。发现交联结构形成了多孔材料，而仅由弱相互作用稳定的材料。所有测试的肽对所有测试的细胞系均无细胞毒性。所选肽也没有遗传毒性，也没有诱导免疫系统反应。我们将继续研究基于 IV 型胶原片段的多孔材料的应用，这些材料能够形成稳定的空间结构，作为再生医学中有用的支架。

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