Promoting Human Intestinal Organoid Formation and Stimulation Using Piezoelectric Nanofiber Matrices

Abstract

Human organoid model systems have changed the landscape of developmental biology and basic science. They serve as a great tool for human-specific interrogation. In order to advance our organoid technology, we aimed to test the compatibility of a piezoelectric material with organoid generation, because it will create a new platform with the potential for sensing and actuating organoids in physiologically relevant ways. We differentiated human pluripotent stem cells into spheroids following the traditional human intestinal organoid (HIO) protocol atop a piezoelectric nanofiber scaffold. We observed that exposure to the biocompatible piezoelectric nanofibers promoted spheroid morphology 3 days sooner than with the conventional methodology. At day 28 of culture, HIOs grown on the scaffold appeared similar. Both groups were readily transplantable and developed well-organized laminated structures. Graft sizes between groups were similar. Upon characterizing the tissue further, we found no detrimental effects of the piezoelectric nanofibers on intestinal patterning or maturation. Furthermore, to test the practical feasibility of the material, HIOs were also matured on the nanofiber scaffolds and treated with ultrasound, which lead to increased cellular proliferation which is critical for organoid development and tissue maintenance. This study establishes a proof of concept for integrating piezoelectric materials as a customizable platform for on-demand electrical stimulation of cells using remote ultrasonic waveforms in regenerative medicine.

Impact Statement

This study introduces an electrospun piezoelectric nanofiber scaffold into the differentiation protocol of human intestinal organoids. The integration of this material successfully enhances tissue patterning and spheroid morphogenesis, producing transplantable organoids comparable to conventional methods. When activated with low intensity pulsed ultrasound, the scaffold further promotes organoid proliferation. Importantly, the study demonstrates that PVDF TrFE is biocompatible with key stages of intestinal organoid generation, that its hydrophobic nature supports morphogenesis and structural organization, and that it establishes a novel platform to investigate the effects of electrical stimulation on organoid development and function, advancing organoid bioengineering and regenerative applications.