Kim Suryeon, Oh Inhwan, Kim Juhyun, Lee Nayeong, Go Jun, Jin Yang, Kim Won June, Lee Heedoo
Department of Biology and Chemistry, Changwon National University, Changwon, 51140, Republic of Korea.
Laboratory Animal Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea.
Sci Rep. 2025 Jul 12;15(1):25261. doi: 10.1038/s41598-025-10050-w.
Extracellular vesicles (EVs) are nano-sized particles released by various cell types that facilitate intercellular communication by transferring bioactive molecules. Owing to their biocompatibility, EVs are widely used as drug delivery vehicles. However, freezing EVs at - 70 °C can induce aggregation, reducing their effective concentration and drug delivery efficiency. In this study, we evaluated the use of water-bath sonication to disperse aggregated EVs and compared its effectiveness with regular pipetting. Frozen EVs showed reduced total concentration and increased aggregation relative to fresh EVs. Sonication at power level 3 (40 kHz, 100 W) significantly increased EV concentration and reduced aggregation. Unlike pipetting, only sonication effectively dispersed aggregated EVs, though subsequent pipetting caused reaggregation. In vivo, aggregated EVs were detected in the bronchoalveolar lavage fluid of mice treated with frozen EVs, whereas mice receiving sonicated EVs exhibited fewer aggregates and enhanced cellular uptake. Molecular dynamics simulations supported the effectiveness of sonication in dispersing EVs. In conclusion, water-bath sonication is a simple and effective method to restore the functionality of freeze-thawed EVs, improving their intracellular delivery and therapeutic potential.
细胞外囊泡（EVs）是由多种细胞类型释放的纳米级颗粒，通过传递生物活性分子促进细胞间通讯。由于其生物相容性，EVs被广泛用作药物递送载体。然而，在-70°C下冷冻EVs会诱导聚集，降低其有效浓度和药物递送效率。在本研究中，我们评估了使用水浴超声处理来分散聚集的EVs，并将其有效性与常规移液进行比较。相对于新鲜的EVs，冷冻的EVs显示出总浓度降低和聚集增加。功率水平为3（40kHz，100W）的超声处理显著提高了EV浓度并减少了聚集。与移液不同，只有超声处理有效地分散了聚集的EVs，尽管随后的移液会导致重新聚集。在体内，在用冷冻的EVs处理的小鼠的支气管肺泡灌洗液中检测到聚集的EVs，而接受超声处理的EVs的小鼠表现出较少的聚集物并增强了细胞摄取。分子动力学模拟支持超声处理在分散EVs方面的有效性。总之，水浴超声处理是一种简单有效的方法，可恢复冻融后EVs的功能，提高其细胞内递送和治疗潜力。
