Zhang Anyu, van Genderen Anne Metje, Liu Bingyan, Qian Junyi, Iamsamang Jirawat, Wang Ziyu, Castilho Miguel, Akhavan Behnam
School of Biomedical Engineering, University of Sydney, Sydney, Australia.
School of Physics, University of Sydney, Sydney, Australia.
Mater Today Bio. 2025 May 29;33:101923. doi: 10.1016/j.mtbio.2025.101923. eCollection 2025 Aug.
Melt electrowriting (MEW) enables the fabrication of highly controlled, open-pore tubular constructs for replicating the complex architectures of vascular, renal, and other tissues. However, a key challenge is to functionalize their surfaces so that they not only support but also instruct key biological interactions, particularly in promoting vascularization. Here, we propose plasma immersion ion implantation (PIII) as a biofunctionalization strategy for open-pore tubular constructs fabricated by MEW. Surface chemistry analysis confirmed homogeneous treatment across PIII-treated MEW 3D structures, while uniaxial tensile tests demonstrated no significant changes in mechanical properties following the treatment. Electron paramagnetic resonance (EPR) data provided evidence of the formation of a stable, radical-rich surface, which was further validated by fluorescence imaging with a model molecule, confirming the radicals' role in enabling uniform covalent biomolecule attachment. The PIII-treated MEW constructs were covalently functionalized with vascular endothelial growth factor (VEGF), thereby modulating the behavior of seeded cells. Endothelialization studies using conditionally immortalized glomerular endothelial cells (ciGEnC) demonstrated that VEGF-immobilized MEW tubes effectively support monolayer formation, achieving outcomes comparable to those observed with VEGF supplementation in culture media. Remarkably, the immobilized VEGF sustained endothelialization with a similar effectiveness to traditional VEGF suspension methods over prolonged culture conditions (21 days), but without the need for continuous VEGF supplementation. These findings establish a novel biofunctionalization strategy for vascularized tissue engineering scaffolds and pave the way for plasma-modified MEW tubes as platforms for preclinical models and regenerative medicine applications.
熔体电写(MEW)能够制造高度可控的开孔管状结构,用于复制血管、肾脏和其他组织的复杂结构。然而,一个关键挑战是使其表面功能化,以便它们不仅能支持而且能引导关键的生物相互作用,特别是在促进血管生成方面。在这里,我们提出等离子体浸没离子注入(PIII)作为一种用于由MEW制造的开孔管状结构的生物功能化策略。表面化学分析证实了对经PIII处理的MEW三维结构进行了均匀处理,而单轴拉伸试验表明处理后力学性能没有显著变化。电子顺磁共振(EPR)数据提供了形成稳定的、富含自由基表面的证据,这通过与模型分子的荧光成像进一步得到验证,证实了自由基在实现均匀共价生物分子附着中的作用。经PIII处理的MEW结构用血管内皮生长因子(VEGF)进行了共价功能化,从而调节了接种细胞的行为。使用条件永生化肾小球内皮细胞(ciGEnC)进行的内皮化研究表明,固定有VEGF的MEW管有效地支持单层形成,取得的结果与在培养基中添加VEGF所观察到的结果相当。值得注意的是,在延长的培养条件(21天)下,固定化的VEGF以与传统VEGF悬浮方法相似的有效性维持内皮化,但无需持续补充VEGF。这些发现为血管化组织工程支架建立了一种新的生物功能化策略,并为作为临床前模型和再生医学应用平台的等离子体改性MEW管铺平了道路。