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3D printing of microscopic bacterial communities
Jodi L Connell1, Eric T Ritschdorff, Marvin Whiteley
1Departments of Chemistry and Biochemistry and Molecular Genetics and Microbiology, and Institute of Cell and Molecular Biology, The University of Texas at Austin, Austin, TX 78712.
Summary
Microscopic 3D printing creates bacterial communities in any shape. This novel method reveals how Pseudomonas aeruginosa protects Staphylococcus aureus from antibiotics.
Area of Science:
- Microbiology
- Biotechnology
- Biofabrication
Background:
- Bacteria use cell-cell communication for adaptive phenotypes, but in vitro studies average properties over large populations.
- Key molecular determinants of bacterial fitness and pathogenicity vary at micrometer scales within dense cellular aggregates.
- Understanding cell-cell interactions in complex environments requires more relevant cellular models.
Purpose of the Study:
- To develop a novel microscopic 3D printing strategy for organizing multiple bacterial populations in defined 3D geometries.
- To create more relevant cellular models for assessing bacterial phenotypes and interactions in complex environments.
- To investigate how bacterial community structure influences antibiotic resistance.
Main Methods:
- A laser-based lithographic technique using focal cross-linking of polypeptide molecules in gelatin to form microscopic containers.
- Bacteria suspended in gelatin are localized within sealed, porous cavities supporting growth and diffusion of biologically active species.
- Construction of co-cultures, including nested and adjacent colonies, and assessment of antibiotic resistance.
Main Results:
- The 3D printing strategy successfully organized multiple bacterial populations into various 3D geometries, including adjacent, nested, and free-floating colonies.
- The porous gelatin material supported bacterial growth and allowed diffusion of signaling molecules and antibiotics.
- A picoliter-volume aggregate of Staphylococcus aureus exhibited significant resistance to β-lactam antibiotics when enclosed by a Pseudomonas aeruginosa shell.
Conclusions:
- Microscopic 3D printing offers unprecedented control for constructing complex bacterial communities.
- This technique enables the study of microscale bacterial interactions and their impact on phenotypes like antibiotic resistance.
- Bacterial community architecture, formed via 3D printing, can significantly influence pathogen survival and treatment efficacy.
