Takeda Nobutaka, Tsuchiya Atsunori, Mito Masaki, Natsui Kazuki, Natusi Yui, Koseki Yohei, Tomiyoshi Kei, Yamazaki Fusako, Yoshida Yuki, Abe Hiroyuki, Sano Masayuki, Kido Taketomo, Yoshioka Yusuke, Kikuta Junichi, Itoh Tohru, Nishimura Ken, Ishii Masaru, Ochiya Takahiro, Miyajima Atsushi, Terai Shuji
Division of Gastroenterology and Hepatology, Graduate School of Medical and Dental Sciences, Niigata University, 1-757 Asahimachi-Dori, Chuo-Ku, Niigata, 951-8510, Japan.
Future Medical Research Center for Exosome and Designer Cell (F-EDC), Niigata University, 1-757 Asahimachi-Dori, Chuo-Ku, Niigata, 951-8510, Japan.
Inflamm Regen. 2023 Oct 9;43(1):48. doi: 10.1186/s41232-023-00299-x.
The progression of liver fibrosis leads to portal hypertension and liver dysfunction. However, no antifibrotic agents have been approved for cirrhosis to date, making them an unmet medical need. Small extracellular vesicles (sEVs) of mesenchymal stem cells (MSCs) are among these candidate agents. In this study, we investigated the effects of sEVs of MSCs, analyzed their distribution in the liver post-administration, whether their effect was dose-dependent, and whether it was possible to collect a large number of sEVs.
sEVs expressing tdTomato were generated, and their uptake into constituent liver cells was observed in vitro, as well as their sites of uptake and cells in the liver using a mouse model of liver cirrhosis. The efficiency of sEV collection using tangential flow filtration (TFF) and changes in the therapeutic effects of sEVs in a volume-dependent manner were examined.
The sEVs of MSCs accumulated mostly in macrophages in damaged areas of the liver. In addition, the therapeutic effect of sEVs was not necessarily dose-dependent, and it reached a plateau when the dosage exceeded a certain level. Furthermore, although ultracentrifugation was commonly used to collect sEVs for research purposes, we verified that TFF could be used for efficient sEV collection and that their effectiveness is not reduced.
In this study, we identified some unknown aspects regarding the dynamics, collection, and capacity dependence of sEVs. Our results provide important fundamentals for the development of therapies using sEVs and hold potential implications for the therapeutic applications of sEV-based therapies for liver cirrhosis.
肝纤维化进展会导致门静脉高压和肝功能障碍。然而，迄今为止尚无抗纤维化药物被批准用于肝硬化治疗，这使其成为未满足的医疗需求。间充质干细胞（MSC）的小细胞外囊泡（sEV）是这些候选药物之一。在本研究中，我们调查了MSC的sEV的作用，分析了给药后它们在肝脏中的分布、其作用是否呈剂量依赖性以及是否有可能大量收集sEV。
生成表达tdTomato的sEV，在体外观察其被肝实质细胞摄取的情况，并使用肝硬化小鼠模型观察其在肝脏中的摄取部位和细胞。研究了使用切向流过滤（TFF）收集sEV的效率以及sEV治疗效果随体积的变化情况。
MSC的sEV主要积聚在肝脏受损区域的巨噬细胞中。此外，sEV的治疗效果不一定呈剂量依赖性，当剂量超过一定水平时达到平台期。此外，尽管超速离心通常用于研究目的收集sEV，但我们证实TFF可用于高效收集sEV且其有效性不会降低。
在本研究中，我们确定了关于sEV的动力学、收集和容量依赖性的一些未知方面。我们的结果为开发使用sEV的疗法提供了重要基础，并对基于sEV的肝硬化治疗应用具有潜在意义。
