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中华细胞与干细胞杂志(电子版)

中华细胞与干细胞杂志(电子版) ›› 2023, Vol. 13 ›› Issue (06) : 363 -369. doi: 10.3877/cma.j.issn.2095-1221.2023.06.006

综述

工程化小细胞外囊泡的制备方法和应用前景
付爽1, 刘语菲1, 周洁1, 周振宇2, 王淑芳1,†()   
  1. 1. 300071　天津，南开大学生命科学学院生物活性材料教育部重点实验室
    2. 250031　济南，中国人民解放军联勤保障部队第九六〇医院骨科
  • 收稿日期:2023-08-25 出版日期:2023-12-01
  • 通信作者: 王淑芳
  • 基金资助:
    科技部重大专项(2020YFA0803701); 天津市科技计划项目(22JCZDJC00470); 济南市临床医学科技创新计划(202225066)

Preparation method and applicative prospect of engineered sEVs

Shuang Fu1, Yufei Liu1, Jie Zhou1, Zhenyu Zhou2, Shufang Wang1,†()   

  1. 1. Key Laboratory of Bioactive Materials for the Ministry of Education College of Life Sciences, Nankai University, Tianjin 300071, China
    2. Department of Orthopedics, the 960th Hospital of the PLA Joint Logistics Support Force, Jinan 250031, China
  • Received:2023-08-25 Published:2023-12-01
  • Corresponding author: Shufang Wang
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引用本文：

付爽, 刘语菲, 周洁, 周振宇, 王淑芳. 工程化小细胞外囊泡的制备方法和应用前景[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(06): 363-369.

Shuang Fu, Yufei Liu, Jie Zhou, Zhenyu Zhou, Shufang Wang. Preparation method and applicative prospect of engineered sEVs[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2023, 13(06): 363-369.

小细胞外囊泡（sEVs）凭借优异的生物相容性、固有稳定性以及低免疫原性，被认为是理想的天然纳米药物递送系统。基于天然sEVs的局限性，工程化sEVs的构建旨在通过负载药物等手段提高其内容物质量和靶向特异性，使sEVs具有更强的药物递送效率，从而最大限度地提高治疗效果。当前的研究已经证实工程化sEVs在生物医学领域和临床应用上表现出极大的潜力。本文重点综述了国内外工程化sEVs制备策略的最新研究成果及其应用前景，针对亲本细胞和sEVs的直接修饰方法进行总结分类并通过举例比较不同方法之间的优缺点；阐述了工程化sEVs在肿瘤、神经系统疾病和组织再生领域的研究进展；评估现阶段工程化sEVs在制备过程中所面临的问题和未来的发展机遇。

关键词: 小细胞外囊泡, 载药, 工程化小细胞外囊泡, 药物递送

Small extracellular vesicles (sEVs) have been considered an ideal natural nano-drug delivery system due to their excellent biocompatibility, inherent stability and low immunogenicity. Based on natural sEVs' limitations, engineered sEVs' construction aims to improve the quality of their contents and target specificity by loading drugs so that sEVs have stronger drug delivery efficiency, thereby maximizing the therapeutic effect. Current studies have confirmed that engineered sEVs have great potential in the biomedical field and clinical applications. This paper focuses on the latest research results and application prospects of the preparation strategies of engineered sEVs at home and abroad. The direct modification methods of parental cells and sEVs were summarized and classified, and the advantages and disadvantages of different methods were compared by examples. The research progress of engineered sEVs of tumors, nervous system diseases and tissue regeneration is described. The problems faced in the preparation of engineered sEVs at the present stage and their development opportunities in the future are evaluated.

Key words: Small extracellular vesicles, Drug loading, Engineered small extracellular vesicles, Drug delivery
图1 EVs的生物学功能[17]注：EVs携带多种生物分子，如核酸（DNA和RNA）、蛋白质（HSP70、HSP90、ALIX和TSG101）、氨基酸和代谢物等。此外，还有一些跨膜蛋白（CD81、CD9、CD63、Flotillin-1）和脂质（胆固醇）。基于这些活性物质，细胞可通过EVs进行细胞间传递信息和物质交换，从而实现多种生物学功能如调控基因转录和翻译、细胞的存活和增殖、血管生成和伤口愈合、废物管理、宿主-微生物相互作用与病毒免疫、平衡免疫应答，调节中枢和外周免疫、受体-配体信号转导、细胞凋亡、细胞分化和瘤变、细胞迁移和转移性疾病、代谢重编程和调节等
图2 EVs在组织再生领域的工程化设计策略[20]注：a图为通过慢病毒转染或电穿孔对亲本细胞进行基因修饰，以上调某些内容物的分泌（miRNAs、细胞因子等）。因此，衍生的EVs将携带可促进组织修复的特定内容物。b图为外部刺激（缺氧、添加NO、促炎性刺激等）作用于亲本细胞也会对分泌的EVs产生间接影响。将供体细胞预分化培养成定向谱系也可提高EVs的再生潜能。c图为EVs逐层纯化的工程化思路。首先，从EVs混合物中分离外泌体，然后确认在组织工程中具有更强功能的特定内容物。d图为利用3D培养和生物反应器优化亲本细胞的培养环境。可以通过监测和控制培养参数来提高EVs的可扩展性和一致性
图3 EV s在组织工程中的应用[61]注：EVs在组织工程中的给药方式包括注射给药、鼻腔给药以及口服给药等。在心脏再生领域中，间充质干细胞来源的EVs被广泛应用，如心脏贴片。同时在肾脏疾病中，EVs也是有潜力的治疗策略。在中枢神经系统疾病中，由于EVs可跨越血脑屏障，EVs作为药物递送系统用于疾病的治疗。此外，在牙髓再生领域，主要用于治疗的是人牙髓间充质干细胞来源的EVs。在骨再生和软骨再生领域中，EVs通过携带的蛋白质和miRNA介导细胞间通讯，从而发挥抗炎和保护软骨等作用
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