新型手性自组装短肽R-LIFE-1的理化性质及其对外泌体的控释研究

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

本研究旨在探究手性自组装短肽R-LIFE-1对外泌体的包裹控释作用。采用原子力显微镜、透射电镜及冷冻扫描电镜等观察短肽R-LIFE-1的成胶能力及形态结构;采用光镜观察及荧光染色探究R-LIFE-1的生物相容性;采用超滤离心法提取外泌体并采用Western blot、纳米粒径追踪分析(NTA)、透射电镜检测外泌体质量;采用激光共聚焦显微镜及BCA蛋白定量探究短肽水凝胶对外泌体的控释效果。结果显示手性自组装短肽R-LIFE-1能够在离子触发下快速自组装形成稳定的纳米纤维网络膜片状结构,具有良好的生物相容性,能够负载外泌体对它进行包裹控释,提高外泌体的利用效率。本研究证明短肽R-LIFE-1可对外泌体进行控释,是一种理想的组织工程材料。

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 selfassembling 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 crosslinked,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.