3D bioprinting of colon organoids in ultrashort self-assembling and decorated peptide matrices

Research into bioinks for organoid culture has the potential to revolutionize our understanding of organ development, function, and disease. Organoids are three-dimensional (3D) cultures of source tissue grown in a support matrix and specialized media. However, the use of animal-derived matrices has limited the potential of organoids in research and therapy. To overcome this limitation, researchers have turned to biofunctional synthetic hydrogel networks to reproduce parameters that govern organoid formation. This study aims to investigate RGD- and YIGSR-decorated ultrashort self-assembling peptides as a modular synthetic hydrogel for organoid culture and 3D bioprinting. Using these motifs, we derived fibronectin (FIB)- and laminin (LAM)-decorated peptides, which self-assemble into nanofibrous hydrogels. We assessed the physicochemical properties of various peptide mixtures. Our findings confirmed the biocompatibility of these formulations and their organoid-forming potential. Subsequently, we identified the most effective scaffolds for organoid formation. We assessed the polarity, differentiation, and functionality of organoids cultured within these scaffolds. We also characterized the properties of a bioprinted construct. This study identifies two formulations, FIB (low) and LAM (high), that favor cell polarization within the cultured organoids as early as day 4. Moreover, these scaffolds were able to induce a gene expression profile resembling the organoids cultured in Matrigel. These peptides were also demonstrated to be suitable for bioprinting at various concentrations without compromising cell viability. Overall, this study demonstrates the promise of modular RGD- and YIGSR-decorated ultrashort self-assembling peptides as effective synthetic hydrogels for organoid culture and 3D bioprinting. These biofunctional peptides provide scaffold effectiveness for advanced organoid manipulation.