From Healthy to Cancerous: A Bioengineered 3D Intestinal Model Using High-Resolution DLP Bioprinting of PEGDA-Based Bioinks

The intestinal epithelium is a dynamic interface essential for nutrient absorption, immune communication, and barrier integrity. Disruption of this system plays a pivotal role in diseases such as colorectal cancer (CRC), a leading cause of cancer-related mortality. Yet, most in vitro models lack the native villi-crypt architecture, limiting their physiological relevance for studying epithelial homeostasis and oncogenic transformation [1,2].

We hypothesize that a photopolymerizable, bioprinted 3D intestinal construct mimicking gut microanatomy can enable more accurate in vitro modeling of healthy and cancerous states. To this end, we designed a biomimetic scaffold via CAD and fabricated it using high-resolution (50 µm) digital light processing (DLP) bioprinting with PEGDA-based bioinks. Parameters were optimized (20 mW/cm², 3 s exposure) to ensure geometric fidelity. Constructs were PBS-washed for seven days and functionalized with poly-L-lysine to enhance cellular adhesion.

Scanning electron microscopy confirmed architectural replication, with villi heights of ~494 µm, 629 µm, and 779 µm (Figure 1). Caco-2 cells were cultured for 21 days to induce epithelial differentiation. Barrier function is monitored via TEER, and gene/protein profiling is underway to assess epithelial homeostasis and early oncogenic markers.

This tunable, structurally biomimetic platform overcomes limitations of current systems, offering a promising tool for disease modeling, mechanistic studies, and high-content drug screening.

References:

[1] Nikolaev, M. et al. Nature 2020, 585, 574–578. doi:10.1038/s41586-020-2724-8

[2] Ballerini, M. et al. Nat. Biomed. Eng 2025. doi:10.1038/s41551-024-01318-z

Acknowledgments:

This research acknowledges the funding provided by UID/50026 (3B’s Research Group, University of Minho), CAN-TARGET (NORTE2030-FEDER-02705300, CCDR-N), and the European Commission through ONCOSCREEN (ID: 101097036) and EngVIPO (ID: 101183041) projects.