Department of Biomedical Engineering, School of Medical Technology, Beijing Institute of Technology, Beijing 100081, China.
Eye Center, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing 102218, China.
Lab Chip. 2024 Mar 26;24(7):1957-1964. doi: 10.1039/d3lc01014g.
Electroporation (in which the permeability of a cell membrane is increased transiently by exposure to an appropriate electric field) has exhibited great potential of becoming an alternative to adeno-associated virus (AAV)-based retina gene delivery. Electroporation eliminates the safety concerns of employing exogenous viruses and exceeds the limit of AAV cargo size. Unfortunately, several concerns (, relatively high electroporation voltage, poor surgical operability and a lack of spatial selectivity of retina tissue) have prevented electroporation from being approved for clinical application (or even clinical trials). In this study, a flexible micro-electrode array for retina electroporation (FERE) was developed for retina electroporation. A suitably shaped flexible substrate and well-placed micro-electrodes were designed to adapt to the retina curvature and generate an evenly distributed electric field on the retina with a significantly reduced electroporation voltage of 5 V. The FERE provided (for the first time) a capability of controlled gene delivery to the different structural layers of retina tissue by precise control of the distribution of the electrical field. After ensuring the surgical operability of the FERE on rabbit eyeballs, the FERE was verified to be capable of transfecting different layers of retina tissue with satisfactory efficiency and minimum damage. Our method bridges the technical gap between laboratory validation and clinical use of retina electroporation.
电穿孔（通过暴露于适当的电场来暂时增加细胞膜的通透性）在替代腺相关病毒（AAV）为基础的视网膜基因传递方面表现出巨大的潜力。电穿孔消除了使用外源病毒的安全问题，并超过了 AAV 有效载荷大小的限制。不幸的是，一些问题（相对较高的电穿孔电压、较差的手术操作性和视网膜组织缺乏空间选择性）阻碍了电穿孔被批准用于临床应用（甚至临床试验）。在这项研究中，开发了一种用于视网膜电穿孔的柔性微电极阵列（FERE）。设计了适当形状的柔性基底和位置良好的微电极，以适应视网膜的曲率，并在视网膜上产生均匀分布的电场，同时显著降低电穿孔电压至 5V。FERE 通过精确控制电场的分布，首次提供了对视网膜组织不同结构层进行受控基因传递的能力。在确保 FERE 在兔眼球上的手术操作性之后，验证了 FERE 能够以令人满意的效率和最小的损伤转染不同的视网膜组织层。我们的方法弥合了视网膜电穿孔在实验室验证和临床应用之间的技术差距。
