A high-throughput gut-on-chip platform to study the epithelial responses to enterotoxins

Area of Science:

• Gastroenterology
• Toxicology
• Cell Biology

Background:

• Enterotoxins pose significant risks to food safety and human health, necessitating effective research models.
• Existing methods for studying enterotoxin effects on the intestinal epithelium are limited in throughput, robustness, and translatability.
• There is a critical need for advanced platforms to investigate toxin-specific epithelial damage and understand enterotoxin contamination and clinical applications.

Purpose of the Study:

• To develop and validate a Caco-2 tubular model for studying enterotoxin impacts on the human intestinal epithelium.
• To characterize distinct pathogenic mechanisms of various enterotoxins using this novel model.
• To provide a high-throughput, multi-parametric tool for enterotoxin research.

Main Methods:

• Caco-2 tubules were exposed to enterotoxins (nigericin, ochratoxin A, patulin, melittin).
• Transepithelial electrical resistance (TEER) was measured to assess barrier permeability.
• Lactate dehydrogenase (LDH) release assays and DRAQ7 staining evaluated cytotoxicity.
• Actin cytoskeleton was analyzed to observe changes in cell morphology.

Main Results:

• Dose-dependent reductions in barrier permeability (TEER) were observed with higher sensitivity than conventional models.
• Cytotoxicity was detected only after exposure to melittin and ochratoxin A.
• Nigericin specifically induced changes in cell morphology via actin cytoskeleton alterations.

Conclusions:

• The Caco-2 tubular model accurately reflects distinct enterotoxin pathogenic mechanisms.
• This model offers enhanced sensitivity for detecting barrier function changes and evaluating cytotoxicity.
• The Caco-2 tubular system is a promising multi-parametric, high-throughput tool bridging current research and in vivo models.