Thermoforming for Small Feature Replication in Melt Electrowritten Membranes to Model Kidney Proximal Tubule

Abstract

A novel approach merging melt electrowriting (MEW) with matched die thermoforming to achieve scaffolds with micron-sized curvatures (200 – 800 µm versus 1000 µm of mandrel printing) for in vitro modeling of the kidney proximal tubule (PT) is proposed. Recent advances in this field emphasize the relevance of accurately replicating the intricate tissue microenvironment, particularly the curvature of the nephrons’ tubular segments. While MEW offers promising capabilities for fabricating highly and porous precise 3D structures mimicking the PT, challenges persist in approximating the diameter of tubular scaffolds to match the actual PT. The thermoformed MEW membranes retain the initial MEW printing design parameters (rhombus geometry, porosity > 45%) while accurately following the imprinted curvature (ratios between 0.67-0.95). PT epithelial cells cultured on these membranes demonstrate the ability to fill in the large pores of the membrane by secreting their own collagen IV-rich extracellular matrix and form an organized, functional, and tight monolayer expressing characteristic PT markers. Besides approximating PT architecture, this setup maximizes the usable surface area for cell culture and molecular readouts. By closely mimicking the structural intricacies of native tissue architecture, this approach enhances the biomimetic fidelity of engineered scaffolds, offering potential applications beyond kidney tissue engineering.