Intestinal Villi Model with Blood Capillaries Fabricated Using Collagen-Based Bioink and Dual-Cell-Printing Process

The human intestine, a vital organ in our digestive system, shows an anatomically complex architecture. The fabrication of three-dimensional (3D) intestinal models containing villus structures has been an important topic for intestine regeneration or organ-on-a-chip, because a 3D model can provide broad surface area and help absorption and transportation of digested nutrients. In this study, we developed a 3D intestinal villi model containing an epithelium layer and a blood capillary structure, using an innovative cell-printing process. The epithelium and capillary network of the 3D model were fabricated using two collagen-based bioinks laden with Caco-2 cells and human umbilical vein endothelial cells (HUVECs). The fabricating conditions were optimized to obtain a unique 3D villus structure, with capillary in the core and high cell viability. A fabricated single villus was 183 ± 12 μm in diameter and 770 ± 42 μm in height, which means the aspect ratio of the structure was 4.2 ± 0.3. The results indicate that the cell-laden intestinal villi successfully mimicked the 3D geometry of human intestinal villi. In vitro cellular activity of the 3D villi model containing epithelium and capillary demonstrated significantly higher cell growth and expression of enzymes and MUC17, compared to those of 2D models and a 3D villi model without the capillary network. The suggested 3D intestinal villi also exhibited the enhancement of the barrier function as compared to those of the others, and even demonstrated an increase of the permeability coefficient of FITC-dextran and glucose uptake ability (FITC, fluorescein isothiocyanate). These results indicate that a 3D intestinal villi model would be a highly promising for mimicking the human intestine.