Guha Ray Preetam, Rajasekaran Ragavi, Pratihar Bitan, De Sirshendu, Dhara Santanu, Fussenegger Martin
ETH Zurich, Department of Biosystems Science and Engineering, Klingelbergstrasse 48, Basel, CH-4056, Switzerland.
Biomaterials and Tissue Engineering Laboratory, School of Medical Science and Technology (SMST), Indian Institute of Technology Kharagpur, Kharagpur, 721302, India.
Adv Sci (Weinh). 2025 Mar;12(9):e2412257. doi: 10.1002/advs.202412257. Epub 2025 Jan 10.
Neo-vascularization plays a key role in achieving long-term viability of engineered cells contained in medical implants used in precision medicine. Moreover, strategies to promote neo-vascularization around medical implants may also be useful to promote the healing of deep wounds. In this context, a biocompatible, electroconductive borophene-poly(ε-caprolactone) (PCL) 3D platform is developed, which is called VOLT, to support designer cells engineered with a direct-current (DC) voltage-controlled gene circuit that drives secretion of vascular endothelial growth factor A (VEGFA). The VOLT platform consists of a 3D-printed borophene-PCL honeycomb-shaped matrix decorated with borophene-PCL nanofibers by electrospinning. The honeycomb structure provides mechanical stability, while the nanofibers facilitate the adhesion, migration, and proliferation of the engineered cells. The cells incorporate a DC-powered reactive oxygen species (ROS)-sensing gene circuit wired to an engineered synthetic promoter that triggers secretion of VEGFA to promote vascularization in the adjacent extracellular matrix. Cells engineered with this gene circuit and enclosed in the VOLT matrix, termed the VOLT system, can be simply triggered using off-the-shelf AA batteries, utilizing the established ability of a brief DC bias to generate non-cytotoxic levels of ROS. For proof-of-concept, a subcutaneous wound-healing model in rats is chosen. Electrostimulation of a VOLT implant (5 V, 20 s per day) induced secretion of VEGFA, and significantly accelerated neovascularization and granulation tissue formation, resulting in faster wound closure compared with non-stimulated controls. Complete re-epithelialization and dermal regeneration are observed within 15 days of application.
新生血管形成在实现精准医学中使用的医用植入物中所含工程细胞的长期存活方面起着关键作用。此外，促进医用植入物周围新生血管形成的策略可能也有助于促进深度伤口的愈合。在此背景下，开发了一种生物相容性导电硼烯-聚（ε-己内酯）（PCL）3D平台，称为VOLT，以支持用驱动血管内皮生长因子A（VEGFA）分泌的直流（DC）电压控制基因电路工程化的设计细胞。VOLT平台由3D打印的硼烯-PCL蜂窝状基质组成，通过静电纺丝用硼烯-PCL纳米纤维进行装饰。蜂窝结构提供机械稳定性，而纳米纤维促进工程细胞的粘附、迁移和增殖。这些细胞整合了一个由直流供电的活性氧（ROS）传感基因电路，该电路连接到一个工程合成启动子，该启动子触发VEGFA的分泌，以促进相邻细胞外基质中的血管形成。用这种基因电路工程化并封装在VOLT基质中的细胞，称为VOLT系统，可以使用现成的AA电池简单地触发，利用短暂直流偏置产生非细胞毒性水平ROS的既定能力。为了进行概念验证，选择了大鼠皮下伤口愈合模型。对VOLT植入物进行电刺激（5 V，每天20秒）可诱导VEGFA的分泌，并显著加速新生血管形成和肉芽组织形成，与未刺激的对照组相比，伤口闭合更快。在应用后的15天内观察到完全重新上皮化和真皮再生。
