Printing of Patterned, Engineered E. coli Biofilms with a Low-Cost 3D Printer.

Area of Science:

• Biomaterials Engineering
• Synthetic Biology
• Microbiology

Background:

• Biofilms exhibit significant heterogeneity, complicating research and limiting applications.
• Standardized, reproducible biofilm models are crucial for advancing biofilm studies and engineering novel materials.

Purpose of the Study:

• To develop a method for producing standardized, reproducible, and patterned biofilm-inspired materials.
• To combine 3D printing with genetic engineering for creating living, functional biomaterials.

Main Methods:

• Prototyped a low-cost 3D printer utilizing bioink (bacteria in alginate suspension).
• 3D printed Escherichia coli in specific shapes and layers on a calcium-containing substrate.
• Genetically induced curli fiber formation in printed bacteria to mimic biofilm structures.

Main Results:

• Printed bacteria survived in the matrix for over a week.
• Induced curli formation resulted in stable, patterned structures resistant to matrix-dissolving agents.
• Demonstrated the first patterned, biofilm-inspired living materials using genetic and spatial control.

Conclusions:

• This 3D printing and genetic engineering approach yields reproducible, patterned biofilm-mimicking materials.
• These living materials offer potential applications in water filtration, metal ion sequestration, and civil engineering.
• The developed materials can serve as standardizable models for studying curli-containing biofilms.