Three-dimensional multicellular layer structure: an advanced in vitro model for studying inflammatory bowel diseases.

Area of Science:

• Biomedical Engineering
• Gastroenterology
• Regenerative Medicine

Background:

• Inflammatory Bowel Disease (IBD) is a chronic gastrointestinal disorder driven by complex multifactorial causes.
• Extracellular Vesicles (EVs) are emerging as a promising therapeutic strategy for inflammatory conditions due to their anti-inflammatory and regenerative properties.
• Existing 2D cell culture models lack the physiological complexity to accurately study IBD and test therapeutics.

Purpose of the Study:

• To develop a 3D multilayer structure (MLS) model of the intestinal mucosa that mimics native tissue complexity.
• To utilize this MLS model as a platform for evaluating the therapeutic potential of Extracellular Vesicles (EVs) in an Inflammatory Bowel Disease (IBD) context.
• To investigate the interactions between EVs, epithelial cells (Caco-2), and stromal components (fibroblasts) within the 3D model.

Main Methods:

• Constructed a 3D multilayer structure (MLS) model using Caco-2 cells and BJ fibroblasts.
• Induced an Inflammatory Bowel Disease (IBD)-like state in the MLS by stimulating with pro-inflammatory cytokines.
• Administered Extracellular Vesicles (EVs) to the stimulated MLS and analyzed gene expression using RT-qPCR.

Main Results:

• The MLS model successfully integrated Caco-2 cells and fibroblasts, with fibroblasts being essential for Caco-2 cell assembly.
• EV treatment significantly modulated gene expression, increasing the anti-inflammatory gene IL-10 and the stemness gene LGR5.
• The 3D MLS model provided a more accurate representation of tissue architecture and cellular interactions compared to traditional 2D cultures.

Conclusions:

• The developed 3D MLS model serves as a valuable platform for drug testing and studying complex gastrointestinal diseases like IBD.
• Extracellular Vesicles (EVs) demonstrate potential in mitigating inflammation and promoting tissue homeostasis within the IBD model.
• This study highlights the utility of advanced 3D models in regenerative medicine and understanding cellular crosstalk in disease.