3D-Printed Biocompatible Frames for Electrospun Nanofiber Membranes: An Enabling Biofabrication Technology for...

Electrospun nanofiber membranes (ESNFMs) are exceptional biomaterials for tissue engineering, closely mimicking the native extracellular matrix. However, their inherent fragility poses significant handling, processing, and integration challenges, limiting their widespread application in advanced 3D tissue models and biofabricated devices. This study introduces a novel and on-mat framing technique utilizing extrusion-based printing of a UV-curable biocompatible resin (Biotough D90 MF) to create rigid, integrated support structures directly on chitosan-polyethylene oxide (PEO) ESNFMs. We demonstrate fabrication of these circular frames via precise 3D printing and a simpler manual stamping method, achieving robust mechanical stabilization that enables routine laboratory manipulation without membrane damage. The resulting framed ESNFMs maintain structural integrity during subsequent processing and exhibit excellent biocompatibility in standardized extract assays (116.5 ± 12.2% normalized cellular response with optimized processing) and acceptable performance in direct contact evaluations (up to 78.2 ± 32.4% viability in the optimal configuration). Temporal assessment revealed characteristic cellular adaptation dynamics on nanofiber substrates, emphasizing the importance of extended evaluation periods for accurate biocompatibility determination of three-dimensional scaffolds. This innovative biofabrication approach overcomes critical limitations of previous handling methods, transforming delicate ESNFMs into robust, easy-to-use components for reliable integration into complex cell culture applications, barrier tissue models, and engineered systems.