Batista Napotnik Tina, Bello Gianluca, Sinner Eva-Kathrin, Miklavčič Damijan
Faculty of Electrical Engineering, University of Ljubljana, Tržaška 25, 1000, Ljubljana, Slovenia.
Institute of Synthetic Bioarchitectures, University of Natural Resources and Life Sciences (BOKU), Muthgasse 11, 1190, Vienna, Austria.
J Membr Biol. 2017 Oct;250(5):441-453. doi: 10.1007/s00232-017-9968-8. Epub 2017 Jul 22.
Polymersomes, vesicles composed of block copolymers, are promising candidates as membrane alternatives and functional containers, e.g., as potential carriers for functional molecules because of their stability and tunable membrane properties. In the scope of possible use for membrane protein delivery to cells by electrofusion, we investigated the cytotoxicity of such polymersomes as well as the effects of nanosecond electric pulses with variable repetition rate on the shape and permeability of polymersomes in buffers with different conductivities. The polymersomes did not show cytotoxic effects to CHO and B16-F1 cells in vitro in concentrations up to 250 µg/mL (for 48 h) or 1.35 mg/mL (for 60 min), which renders them suitable for interacting with living cells. We observed a significant effect of the pulse repetition rate on electrodeformation of the polymersomes. The electrodeformation was most pronounced in low conductivity buffer, which is favorable for performing electrofusion with cells. However, despite more pronounced deformation at higher pulse repetition rate, the electroporation performance of polymersomes was unaffected and remained in similar ranges both at 10 Hz and 10 kHz. This phenomenon is possibly due to the higher stability and rigidity of polymer vesicles, compared to liposomes, and can serve as an advantage (or disadvantage) depending on the aim in employing polymersomes such as stable membrane alternative architectures or drug vehicles.
聚合物囊泡是由嵌段共聚物组成的囊泡，作为膜替代物和功能容器具有很大潜力，例如，由于其稳定性和可调节的膜性质，可作为功能分子的潜在载体。在通过电融合将膜蛋白递送至细胞的可能应用范围内，我们研究了此类聚合物囊泡的细胞毒性，以及不同重复率的纳秒电脉冲对不同电导率缓冲液中聚合物囊泡形状和渗透性的影响。聚合物囊泡在体外对CHO和B16 - F1细胞在浓度高达250μg/mL（48小时）或1.35mg/mL（60分钟）时未显示出细胞毒性作用，这使其适合与活细胞相互作用。我们观察到脉冲重复率对聚合物囊泡的电极形成有显著影响。电极形成在低电导率缓冲液中最为明显，这有利于与细胞进行电融合。然而，尽管在较高脉冲重复率下变形更明显，但聚合物囊泡的电穿孔性能未受影响，在10Hz和10kHz时均保持在相似范围内。这种现象可能是由于与脂质体相比，聚合物囊泡具有更高的稳定性和刚性，根据使用聚合物囊泡的目的，如稳定的膜替代结构或药物载体，这可能是一个优点（或缺点）。
