Bai Chengmeng, Ren Huifang, Zhang Feng, Li Dandan, Feng Qingyu, Li Ying, Chen Ting, Li Chengxu, Xiao Xuan, Zhang Hengrui, Hu Liang
National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
State Key Laboratory Cultivation Base, Shandong Key Laboratory of Eye Diseases, Eye Institute of Shandong First Medical University, Qingdao, China.
J Nanobiotechnology. 2025 Jul 1;23(1):476. doi: 10.1186/s12951-025-03556-y.
Scalable approaches for enhancing therapeutic small extracellular vesicles (sEVs) production can facilitate the transition of sEVs from bench to bedside and beyond. Here, we present a user-friendly method to manipulate the extracellular mechanical microenvironment of umbilical cord-derived mesenchymal stem cells (MSCs), a promising cell type for generating sEVs with therapeutic benefits, to boost sEV secretion and regenerative bioactivity. The bioreactor system, called the programmable controlled rotating platform (PRP), is designed to apply normal stress on cell surface through centrifugal rotation culture. Experimental analyses suggested that the PRP can promote a 4-fold sEV secretion increase without affecting cell viability and sEV size when compared to the traditional static culture condition. More importantly, PRP-induced MSC-sEVs can significantly promote epithelial cell migration in vitro and accelerate corneal wound healing in a murine model, with suppressed inflammatory responses in wound bed tissue. Further mechanistic investigations revealed that this process involves the activation of cellular transcriptional signals implicated in sEV biogenesis. Concurrently, sEV cargo undergoes remodeling to enrich regenerative and immunoregulatory functions. These findings demonstrate the efficacy of our established platform in advancing sEV production and improving clinical performance, providing a novel sEV-based mechanism for ocular treatments, including corneal epithelialization and even retinal neural regeneration.
增强治疗性小细胞外囊泡（sEVs）产量的可扩展方法能够促进sEVs从实验室到临床应用乃至更广泛领域的转化。在此，我们提出一种用户友好型方法，用于操控脐带间充质干细胞（MSCs）的细胞外机械微环境，这是一种有望产生具有治疗益处的sEVs的细胞类型，以提高sEVs的分泌及再生生物活性。该生物反应器系统称为可编程控制旋转平台（PRP），旨在通过离心旋转培养在细胞表面施加法向应力。实验分析表明，与传统静态培养条件相比，PRP可使sEVs分泌增加4倍，且不影响细胞活力和sEVs大小。更重要的是，PRP诱导的MSC-sEVs可在体外显著促进上皮细胞迁移，并在小鼠模型中加速角膜伤口愈合，同时抑制伤口床组织中的炎症反应。进一步的机制研究表明，这一过程涉及激活与sEV生物发生相关的细胞转录信号。与此同时，sEV的货物发生重塑，以丰富其再生和免疫调节功能。这些发现证明了我们建立的平台在提高sEV产量和改善临床性能方面的有效性，为眼部治疗提供了一种基于sEV的新机制，包括角膜上皮化甚至视网膜神经再生。
