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新型手性自组装肽R-LIFE-1的理化性质及其对外泌体的控释

[Physicochemical properties of a novel chiral self-assembling peptide R-LIFE-1 and its controlled release to exosomes].

作者信息

Luo Xinyi, Su Di, Lu Na, Wan Yuan, Liu Guicen, Luo Zhongli

机构信息

Molecular Medicine and Cancer Research Center, Collage of Basic Medicine, Chongqing Medical University, Chongqing 400016, P. R. China.

出版信息

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2023 Aug 25;40(4):770-777. doi: 10.7507/1001-5515.202207056.

Abstract

This research aims to investigate the encapsulation and controlled release effect of the newly developed self-assembling peptide R-LIFE-1 on exosomes. The gelling ability and morphological structure of the chiral self-assembling peptide (CSAP) hydrogel were examined using advanced imaging techniques, including atomic force microscopy, transmission electron microscopy, and cryo-scanning electron microscopy. The biocompatibility of the CSAP hydrogel was assessed through optical microscopy and fluorescent staining. Exosomes were isolated via ultrafiltration, and their quality was evaluated using Western blot analysis, nanoparticle tracking analysis, and transmission electron microscopy. The controlled release effect of the CSAP hydrogel on exosomes was quantitatively analyzed using laser confocal microscopy and a BCA assay kit. The results revealed that the self-assembling peptide R-LIFE-1 exhibited spontaneous assembly in the presence of various ions, leading to the formation of nanofibers. These nanofibers were cross-linked, giving rise to a robust nanofiber network structure, which further underwent cross-linking to generate a laminated membrane structure. The nanofibers possessed a large surface area, allowing them to encapsulate a substantial number of water molecules, thereby forming a hydrogel material with high water content. This hydrogel served as a stable spatial scaffold and loading matrix for the three-dimensional culture of cells, as well as the encapsulation and controlled release of exosomes. Importantly, R-LIFE-1 demonstrated excellent biocompatibility, preserving the growth of cells and the biological activity of exosomes. It rapidly formed a three-dimensional network scaffold, enabling the stable loading of cells and exosomes, while exhibiting favorable biocompatibility and reduced cytotoxicity. In conclusion, the findings of this study support the notion that R-LIFE-1 holds significant promise as an ideal tissue engineering material for tissue repair applications.

摘要

本研究旨在探究新开发的自组装肽R-LIFE-1对外泌体的包封及控释效果。采用先进成像技术,包括原子力显微镜、透射电子显微镜和低温扫描电子显微镜,研究手性自组装肽(CSAP)水凝胶的凝胶化能力和形态结构。通过光学显微镜和荧光染色评估CSAP水凝胶的生物相容性。通过超滤分离外泌体,并使用蛋白质免疫印迹分析、纳米颗粒跟踪分析和透射电子显微镜评估其质量。利用激光共聚焦显微镜和BCA检测试剂盒对CSAP水凝胶对外泌体的控释效果进行定量分析。结果表明,自组装肽R-LIFE-1在各种离子存在下表现出自发组装,形成纳米纤维。这些纳米纤维相互交联,形成坚固的纳米纤维网络结构,该结构进一步交联形成层状膜结构。纳米纤维具有较大的表面积,能够包封大量水分子,从而形成高含水量的水凝胶材料。这种水凝胶作为细胞三维培养以及外泌体包封和控释的稳定空间支架和负载基质。重要的是,R-LIFE-1表现出优异的生物相容性,保留了细胞的生长和外泌体的生物活性。它能迅速形成三维网络支架,实现细胞和外泌体的稳定负载,同时表现出良好的生物相容性和降低的细胞毒性。总之,本研究结果支持以下观点:R-LIFE-1作为用于组织修复应用的理想组织工程材料具有巨大潜力。

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