3D-Printed Scaffold Mimicking IBD Gut Microenvironments: An In Vitro Model for Bacterial Bioink Growth.

Area of Science:

• Biomaterials Science
• Microbiology
• Gastroenterology

Background:

• Inflammatory bowel disease (IBD) is a chronic gastrointestinal condition associated with gut microbiota dysbiosis.
• Understanding the complex interactions within the gut microbiota is crucial for IBD research.
• Current models for studying gut microbiota may not fully replicate the in vivo environment.

Purpose of the Study:

• To investigate the feasibility of creating a 3D-bioprinted scaffold for culturing Lactococcus lactis.
• To evaluate the suitability of an alginate-agar-soy trypticase bioink for 3D bioprinting.
• To assess the scaffold's capacity to support bacterial viability and growth.

Main Methods:

• Development of a novel bioink composed of alginate, agar, and soy trypticase.
• 3D bioprinting of scaffolds using the developed bioink and Lactococcus lactis.
• Assessment of bioink properties including water absorption and rheology.
• Evaluation of scaffold structural integrity and stability over 24 hours.
• Confocal microscopy to confirm Lactococcus lactis viability and distribution within the scaffold.
• Comparison of bacterial growth (colony-forming units) in 3D scaffolds versus 2D models.

Main Results:

• The alginate-agar-soy trypticase bioink demonstrated excellent water absorption and suitable rheological properties for 3D bioprinting.
• Successfully fabricated robust 3D scaffolds capable of maintaining structural integrity for 24 hours.
• Confocal microscopy confirmed the viability of Lactococcus lactis within the scaffold, showing green fluorescence indicative of live bacteria after 8 hours.
• The 3D scaffold provided a supportive microenvironment, promoting prolonged bacterial survival and proliferation.
• Significantly higher colony-forming units (CFUs) were observed in the 3D scaffold compared to a 2D culture model, indicating enhanced bacterial growth.

Conclusions:

• 3D bioprinting using the developed alginate-agar-soy trypticase bioink is a feasible method for creating bacterial scaffolds.
• The 3D scaffolds offer a stable and supportive environment for Lactococcus lactis, promoting enhanced growth and viability.
• These bacterial scaffolds represent a promising platform for in vitro studies of gut microbiota, particularly for investigating factors influencing microbial communities in IBD and other diseases.